Epidemiological (epizootiological) field surveys are among the primary tasks of the VIRSVB's epizootiological service, which is crucial in the fight against infectious and parasitic diseases of domestic animals.

The tasks of the field epizootiological service are:
– collection of anamnestic and other epidemiological data from the field in the event of a suspected infectious or parasitic disease,
– sampling, field necropsy, or performing rapid field diagnostics,
– assisting competent veterinary organizations and the veterinary inspection in the field with the control of infectious or parasitic diseases 
– providing opinions and recommendations for the purpose of controlling and suppressing diseases.

After each fieldwork, the epizootiological team prepares an epizootiological report. The report is submitted to the client of the epizootiological survey and the Ministry of Agriculture, Forestry and Water Economy of Republika Srpska.

The reason for an epidemiological investigation is a suspicion of an infectious or parasitic disease that exceeds the scale of a sporadic occurrence and the diagnostic capabilities of authorized veterinary organizations.

The VIRSVB's epizootiological teams have controlled many dangerous infectious diseases in the Republika Srpska: anthrax, brucellosis, Q fever, tuberculosis, leptospirosis, salmonellosis, listeriosis, paratuberculosis, pasteurellosis, clostridial infections, enzootic viral pneumonia, avian influenza, rabies, malignant catarrhal fever, bluetongue disease, African swine fever, bovine viral diarrhea, infectious ecthyma, piroplasmosis, etc. Some significant results of the VIRSVB's epizootiological surveys are:
– In early June 2016, a bluetongue outbreak caused by serotype 4 of the virus was located for the first time.
In 2020, a case of rabies in a hunting dog was field-analyzed in Srebrenica for the first time in over 10 years.

In 2023, the epizootiological service in Semberija diagnosed African swine fever for the first time in the Republika Srpska and worked continuously on its containment.

This data confirms that the VIRSVB's epizootiological teams are continuously engaged and ready to respond to combat infectious and parasitic diseases of public interest in the Republika Srpska.

Rabies

Rabies, as a very dangerous acute viral disease of warm-blooded animals and humans, which is transmitted from animals to humans, has always been the focus of work and research at VIRSVB. 
From 1946, experts from the VIRSVB continuously conducted epidemiological and diagnostic investigations, thereby monitoring the situation on the ground. During 1948, the VIRSVB organized a campaign in which over 100,000 dogs were vaccinated against rabies in the territory of Bosnia and Herzegovina. Due to the constant presence of rabies in wild animals, 93 cases of rabies in animals were discovered during 1950. It was recorded that cases of rabies were isolated in two dogs and one ferret in the territories of Banja Luka and Bosanska Dubica (1976), as well as that the Pasteur Institute in Paris determined that the virus isolated from the brain of a horse bitten by a wolf from the territory of Bosanski Grahovo was antigenically identical to the African dog rabies virus (1991). The total number of rabies cases in BiH in the period from 1945 to 1978 was 2,312, of which 1,200 were dogs, 539 cows, 176 sheep, 150 pigs, 93 cats, 77 foxes, 48 horses, and 29 wolves. In recent years, the epidemiological situation regarding rabies has been significantly more favorable. In the period from 1992 to 1994, 40 cases of rabies were confirmed in VIRSVB, and in the period from 1995 to 2001, the total number of positive cases was 205 (in 12 animal species). The percentage of positive animals was highest in foxes (77.56%), followed by cattle (5.85%), dogs (4.39%), and small ruminants (3.90%). Cats accounted for 2.93%, and weasels for 1.95%. Subsequently, in the period up to 2011, there was an average of 37 positive rabies cases annually, with the highest number in foxes (329) and the second-highest in dogs (230). At the end of 2011, oral vaccination of foxes against rabies began, which proved to be an adequate measure, so that in 2012, 7 positive cases of rabies were confirmed: 5 in foxes, 1 in a dog, and 1 in a cow. In Srebrenica, in 2020, the last case of rabies in the Republika Srpska and BiH occurred.

VIRSVB participates in post-vaccination monitoring, which is conducted after the fall fox oral vaccination campaign. Between 2020 and 2024, 530 samples were tested, all of which were negative.

Методе за дијагностику бјеснила које се изводе у ВИРСВБ су сљедеће:
– fluorescent antibody test,
– LISA test (antibodies),
– Real-Time PCR

VIRSVB regularly participates in proficiency tests with the reference laboratory of the World Organisation for Animal Health (WOAH-OIE) and the EU, ANSES (France).

In addition to the Faculty of Veterinary Medicine of the University of Sarajevo, VIRSVB is the only laboratory for the diagnosis of rabies in animals in the area of BiH. As a reminder of VIRSVB's status as the first laboratory in the region to diagnose rabies, its trademark is an emblem with an irregular elliptical shape, in the center of which is a nerve cell with Negri bodies.

Classical swine plague

Classical swine plague is a significant viral disease of domestic and wild pigs that has been known for almost 200 years. It is one of the most significant health and economic diseases of pigs, as it causes enormous losses in production. The significance of this disease is also reflected in international trade, which prohibits the movement of live pigs, pork products, and raw materials for the industry from a country where the disease occurs. Due to its great epizootiological and economic importance, classical swine plague is listed among the particularly dangerous infectious animal diseases.

In Republika Srpska, vaccination against classical swine fever has not been performed since 2020, and the disease was last diagnosed in Republika Srpska and BiH in 2007.

Given the significance of this disease, the Program for the Surveillance of African Swine Fever and Classical Swine Fever in Republika Srpska was adopted in 2019. Through this Program, 8,703 blood and organ samples from domestic and wild pigs from the entire territory of Republika Srpska were submitted to VIRSVB between 2019 and 2022. The samples were analyzed by ELISA for the presence of antibodies and by Real-Time PCR, and no positive reactors were detected.

Also, in the Republika Srpska, in 2023, a Program for Active and Passive Surveillance of Classical and African Swine Fever in the Domestic and Wild Pig Population was adopted. The program was implemented throughout the entire territory of Republika Srpska, and all veterinary organizations were involved, performing sampling and submitting it to the VIRSVB, which is the reference laboratory of Republika Srpska for these two swine infectious diseases. Prior to the emergence of African swine fever, surveillance was conducted in accordance with the planned activities, and after that, analysis for classical swine fever was performed differentially on pigs that were not positive for African swine fever. Accordingly, 159 samples were analyzed using the Real-Time PCR method, with no positive samples. It is significant that the results of the monitoring, in which there were no confirmed positive reactors, show that after the cessation of swine vaccination against classical swine fever, the measures being implemented are yielding good results in controlling this disease.

VIRSVB is the reference laboratory in Republika Srpska and Bosnia and Herzegovina for the diagnosis of classical swine fever.. 

African swine fever

African swine fever is a contagious viral disease of domestic and wild pigs. Wild pigs are considered the main reservoir of the virus in Africa and other areas where the disease is endemic. The disease spreads through contact between infected and healthy pigs, feeding pigs on swill containing contaminated food scraps, contaminated food and water, clothing and footwear, and contaminated items and equipment, vehicles, manure, carcasses of dead and slaughtered animals, rodents, insects.

In recent years, the number of countries where African swine fever appears has increased, and the disease has also been reported in countries in sub-Saharan Africa, Europe, and Asia. African swine fever is a global threat to animal health and food safety, with negative socioeconomic impacts. To control the disease, it is necessary to implement national control programs in cooperation with regional and global coordination. To this end, in 2019, the Program for Surveillance of African Swine Fever and Classical Swine Fever in Republika Srpska was adopted. As the reference laboratory, VIRSVB was responsible for implementing the program, conducting diagnostics, and preparing reports on its implementation. Through this Program, from 2019 to 2022, 8,661 blood and organ samples from domestic and wild pigs from across the entire Republika Srpska were submitted to the VIRSVB. The samples were analyzed by ELISA for the presence of antibodies and by Real-Time PCR, and no positive reactors were detected.

Also, in the Republika Srpska, in 2023, a Program for Active and Passive Surveillance of Classical and African Swine Fever in the Domestic and Wild Pig Population was adopted. The Program was implemented throughout the entire territory of Republika Srpska, and all veterinary organizations were involved, performing sampling and submitting it to the VIRSVB, which is the reference laboratory of Republika Srpska for these two swine infectious diseases. During the implementation of the Program, a case of African swine fever was confirmed, so most activities were focused on measures to combat this disease. The first case of African swine fever in Republika Srpska and BiH was confirmed by VIRSVB on June 22, 2023. After that, The VIRSVB directed its activities toward implementing measures for the early detection, diagnosis, prevention of spread, control, and eradication of the African swine fever infectious disease in accordance with applicable regulations. In the period from June 22 to December 31, 2023, The VRSPB confirmed African swine fever in the Republika Srpska in 24 municipalities on 1,166 outbreak farms, where 49,190 pigs were humanely culled. The average was 41 pigs per infected premises. The VRVSB's activities regarding this disease continued into 2024, when, by the end of July, African swine fever was confirmed in 15 municipalities and 45 outbreaks, of which 2 were in wild boar.

VIRSVB had a large number of activities in connection with the emergence of African swine fever:
– formation of an epizootiological team to identify the outbreak, conduct diagnostics, prevent the spread, and eradicate the disease,
– holding a large number of meetings of the epizootiological team,
– sending samples for confirmation to the EU Reference Laboratory (INIA-CISA, Madrid, Spain), which were confirmed on September 14, 2023,
– conducting educational sessions for hunting ground users in Bijeljina, Šamac, and Gradiška, and preparing proposed measures necessary to control African swine fever in the wild boar population.
– establishment of a molecular diagnostics laboratory in the Bijeljina Business Unit,
– active participation in the media to inform the public, as well as providing recommendations to farmers on procedures and their responsibilities and obligations, all with the aim of preventing the spread and ultimately eradicating the disease,
– holding a meeting with representatives of the Chinese State Veterinary Institute (LVRI, CAAS),
– посјета и одржавање састанка с представницима Свјетске организације за здравље животиња (WOAH-OIE),
– visit and holding a meeting with representatives of the GF-TAD expert committee for Europe,
– Exchange of samples and training of VIRSVB personnel at the Austrian Agency for Health and Food Safety (AGES).

Bird flu

Avian influenza is a viral, highly contagious disease of domestic and wild birds, which has a strong zoonotic potential and poses a major public health threat on a global level. Additionally, this disease has devastating consequences for poultry production, international trade, and the economy. In the past year, it has been detected in wild mammals, and high mortality rates have been recorded in marine mammal populations in South America. According to the latest information from the WHO-OIE, it was detected in dairy cattle in the USA during March and April 2024, which indicates that the virus could cross the species barrier and become established in a new host. In our region, the presence of the avian influenza virus has been confirmed in Serbia, Slovenia, Croatia, and North Macedonia. Given the unfavorable epidemiological situation, surveillance for avian influenza is being conducted in Republika Srpska and BiH.

The first case of avian influenza (H5N1 subtype) in BiH was confirmed in February 2006, in two swans (Cygnus olor) in the area of Jajce. The second case in BiH, and the first case of avian influenza in Republika Srpska, was confirmed in 2017. year, another case of avian influenza was confirmed in Republika Srpska, when a carcass of a domestic chicken from a family farm in a suburb of Prijedor was submitted to VIRSVB, where molecular diagnostics, using the Real-Time PCR method, proved the presence of the causative agent (subtype H5N8). Subsequently, in October 2021. another case of avian influenza was confirmed in Republika Srpska, when a swan carcass found in the bed of the Sava River (Gradiška city) was delivered to the VIRSVB, and the presence of the causative agent (subtype H5N1) was laboratory-confirmed. Also, during this period (2016–2021), the disease was diagnosed in wild birds and poultry in almost all countries in the region and Europe. In February 2024, in Republika Srpska, VIRSVB detected the avian influenza agent (H5N1 subtype) in a population of swans (Cygnus spp.) on the territory of the city of Gradiška.

Saliva and drool

Swinet and foot-and-mouth disease is an acute, highly contagious viral infection, with a high potential for causing significant economic losses in susceptible cloven-hoofed animals. The disease spreads rapidly through cohabitation and contact between infected and susceptible animals, as well as through raw materials and products from infected animals, and by vectors. The disease is present worldwide. This disease poses a major threat to international trade and livestock production due to strict disease control measures and restrictions associated with international trade.

VIRSVB is the reference laboratory in the Republika Srpska and BiH for the diagnosis of rabies. Diagnosis of this disease at VIRSVB is performed using an ELISA test and the Real-Time PCR method.

The mode of virus transmission and its antigenic variability make swine fever and foot-and-mouth disease highly contagious, and the disease is endemic in many parts of the world. There are seven serotypes of the virus (O, A, C, SAT 1, SAT 2, SAT 3, Asia 1), and infection with one serotype does not confer immunity against another. Due to the antigenic diversity of the virus, diagnosis and timely control of the disease are difficult. There is no single laboratory test to confirm the disease, and it is necessary to conduct a combination of several tests to reach a final diagnosis.

The VIRSVB staff has collaborated on and completed the following training in the diagnosis of the oral and paw cavity: 
– Pirbright Institute, the world reference laboratory for salivary gland and foot-and-mouth disease (United Kingdom),
– European Reference Laboratory for Salivary Gland Murrain and Foot-and-Mouth Disease Anses (France),
– Real time training course on Foot and mouth disease EU FMD Kenya
– Internationaler Kurs zur Diagnose von Tierseuchen (Plum Island, New York, USA),
– Exchange of best practice in German, Fridrich Loefler Institute (Riems, Њемачка).

Blue tongue disease

Blue tongue disease, or catarrhal fever of sheep, is a vector-borne viral disease of small ruminants and sometimes cattle. The first case of blue tongue disease serotype 4 in sheep was confirmed by VIRSVB in the village of Vrbnica, Foča, in mid-June 2016. After that, the disease spread rapidly to the eastern part of Republika Srpska, taking on the proportions of an epizootic.

In 2017, VIRSVB conducted an extensive epizootiological survey (mainly in the eastern part of Republika Srpska), where significant data on the movement of the disease were obtained. Specifically, during the 2016 outbreak of bluetongue disease, various forms of the disease were observed in the territory of Republika Srpska. Initially, fever, general weakness, and swelling of the ears, eyelids, and lips may be observed. After the swelling, the next clinical sign, which was noticeable in several sheep, was certainly a copious, frothy discharge from the mouth. Upon opening the mouths of such animals, a swollen and hyperemic tongue with petechiae is observed, and ulcerative changes are visible on the lips. Such changes are visible on the mucous membrane of the lips (the corners of the mouth and the nostrils are covered with ulcerative changes), as well as on the hooves (corium changes, which begin with mild swelling of the joints in the heel region). Changes in the hoof corium are observed in the second stage of the disease. Younger age groups were more severely affected by bluetongue disease. These clinical signs can be considered typical. In the 2017 epidemiological survey, no new cases of the disease were detected in sheep, but according to statements from owners in Pala, Čajniče, Nevesinje, Gacko, Rogatica, and Han Pijesak, abortions and the birth of stillborn lambs were registered in infected sheep. On some farms, some infected sheep did not recover from Bluetongue disease but instead developed a chronic form of the illness.

Based on the epidemiological investigation, the OIE-WAH-2008 Blue Tongue disease was likely spread from the territory of Montenegro, in the border area of the Foča and Gacko epidemiological units. During the primary wave in June and July 2016, the disease spread to western and southern Herzegovina, as well as to the northern part of the eastern region of Republika Srpska. Observing the pattern and timing of the Bluetongue disease outbreak in this epidemiological investigation, it was concluded that the virus, via the vector, further spread in the early fall along the Drina river basin and its tributaries into the Romanija-Podrinje region. Losses were primarily observed in Čajniče (Upper Podrinje). Serological testing and epidemiological data indicate that the mountain ranges were a natural barrier to the spread of the virus, so seroprevalence decreases in Pale, Sokolac, Han Pijesak, and Vlasenica.

In addition to the epidemiological investigation, an entomological study of the presence of culicoids was conducted, yielding the first results on the presence of competent vectors from Sulicoides obsoletus groups and Sulicoides pulicaris комплекса. Ентомолошким испитивањем детектоване су ријетке врсте куликоида у Републици Српској (БиХ), које до тада нису биле присутне.

Ензоотска леукоза говеда

This disease is widespread in many countries on all continents and represents a significant global health problem. It causes significant economic losses and threatens intensive cattle production, primarily due to the condemnation of carcasses with tumorous lesions at the slaughterhouse, as well as reduced milk yield and fertility in infected cattle. The biggest problem in controlling and eradicating this disease is that in most cases, the infection proceeds without visible clinical signs. For this reason, the only way to succeed is to detect infected animals as early as possible and remove them from the herd.

In BiH, a group of scientists in 1984-1985, by agar gel immunodiffusion (AGID) test examined 8,225 cattle on 14 large farms in 1984-1985, and in the process detected the presence of specific antibodies against the bovine enzootic leukosis virus on 5 farms (in 4.7-15.55% of the cattle tested). 
The program of measures on animal health protection in the Republika Srpska defines that all animals older than 24 months, which are on the property at the time of implementation of the measures, must be tested at least once a year, in herds that do not have the status of officially free herds from this disease.

For decades, VIRSVB has been involved in the fight to control and eradicate enzootic bovine leukosis in the Republika Srpska. Testing is primarily performed using an iELISA test, and in a smaller number of cases, an AGID test is also performed. For the period 2020–2024, VIRSVB tested approximately 39,000 bovine serum samples for this disease. After a long period, in the second half of August 2024, the presence of specific antibodies to the bovine enzootic leukosis virus was detected in one of the tested samples.

ВИРСВБ је референтна лабораторија у Републици Српској и БиХ за дијагностику ензоотске леукозе говеда.

Equine Infectious Anemia

Equine infectious anemia is a disease that has been a serious threat to horse breeding worldwide, and in our country as well, for decades. For the past 30 years, the disease has been constantly reported in BiH, and according to certain reports, it was first confirmed in 1952 in a cold-blooded draft horse near Sarajevo, while a series of cases in workhorses at the “Maglic” State Farm in Foča was described shortly thereafter. During 2008–2009, serological monitoring conducted throughout the territory of Republika Srpska proved a prevalence of this disease of 17.76%. This period represents the beginning of the current monitoring of equine infectious anemia in Republika Srpska. In the following years, up to 2023, monitoring was conducted only in municipalities of Republika Srpska where at least 5 cases of equine infectious anemia had been registered in the previous year, and since 2023, all municipalities in Republika Srpska have been included. The municipalities most frequently monitored were: Kotor Varoš, Čelinac, Mrkonjić Grad, Ribnik, and Pale.

Due to the high prevalence of the disease during the 2007–2010 period, a control program for equine infectious anemia was developed, which includes measures such as the tagging (microchipping) of equines, blood collection for serological analysis, and the removal of seropositive animals. The program also includes maintaining a database of tagged and laboratory-tested equines, covering approximately 11,000 animals.

In routine work, VIRSVB analyzes a large number of samples for equine infectious anemia, and in the last four years, that number has been around 2,800 samples. The agar gel immunodiffusion test (AGID), or the Coggins test, is primarily used as the gold standard for diagnosing equine infectious anemia, and the cELISA test is also used in addition to it. In addition to serological tests, in connection with equine infectious anemia, VIRSVB is in constant communication with the field veterinary service and conducts epizootiological surveys.

VIRSVB is the reference laboratory in Republika Srpska and BiH for the diagnosis of equine infectious anemia..

Anthrax

Антракс је древна заразна болест узрокована спорогеном бактеријом Bacillus anthracis. Among the people it is called black anthrax or simply anthrax, thigh rot, evil rot, streaks, etc. It is well known to livestock farmers in certain parts of our country, as it appears continuously in district areas. The districts are, for the most part, karst fields or poorly drained mountain pastures. In the Republika Srpska, anthrax districts include Sokolac (Romanija), Pale, Gacko, Bileća, and the Manjača area.

For decades, VIRSVB has been diagnosing anthrax by isolating and identifying the causative agent from clinical materials, and conducting epidemiological surveys. The last case of anthrax in the Republika Srpska was diagnosed by VIRSVB in 2017 in an unvaccinated lamb from the Sokoc municipality. Additionally, a large number of anthrax cases occurred in Sokoc in 2011, when the disease also appeared in humans.

Brucellosis

Brucellosis is a worldwide prevalent bacterial infectious disease of animals and humans, caused by small Gram-negative cocci of the genus Brucella.

For decades, VIRSVB was the only diagnostic institution in this area and performed all serological tests for brucellosis. From the works of Dr. Vasa Butozan, we learn that after the First World War, all species of Brucella were present throughout Europe, so the area that today is the Republika Srpska was no exception. After World War II, field veterinarians, led by experts from the VIRSVB, fought to suppress and eradicate brucellosis. Thanks to their persistent work in diagnosing brucellosis, as well as that of the entire veterinary service in controlling, suppression and eradication measures, and with the full support of state institutions in controlling animal movement, brucellosis was eradicated from this region in the 1970s. The last case of brucellosis in this area occurred in 1984, when it was detected at the Military Farm in Manjača, near Banja Luka, sheep and people became infected. A rapid response was implemented, during which several thousand sheep from infected flocks, as well as sheep from contact flocks, were humanely culled, along with all other susceptible animals, including dogs and cats.Subsequent annual tests on ruminants revealed no new seropositive animals. In recent history, on the Military Farm in Manjača, near Banja Luka, sheep and people became infected in 1984. A rapid response was implemented, during which several thousand sheep from infected flocks, as well as sheep from contact flocks, were humanely culled, along with all other susceptible animals, including dogs and cats. Subsequent tests conducted annually on ruminants determined that there were no new seropositive animals. In recent history, in the territory of Republika Srpska and BiH, brucellosis caused Brucella melitensis је присутна више од двије деценије. Прво жариште појавило се 2000. године у стаду оваца у Мостару, а 2001. године откривена је код оваца и говеда у Невесињу, и то након што су обољели људи. Већ сљедеће године десила се појава бруцелозе на више локација у Републици Српској и Федерацији БиХ. Услиједиле су године у којима је инциденца бруцелозе сваке године била све већа. Број обољелих људи повећавао се из године у годину, све до 2008. године, када је инциденца бруцелозе достигла свој максимум, јер је само те године откривено 2.007 жаришта, са укупном преваленцијом код малих преживара од 4,55% и 994 обољелих људи. Процијењено је да је на нивоу укупног броја стада, 40% инфицирано бруцелом. У санирању жаришта уништено је преко 70.000 оваца и коза и 260 говеда. Те године ВИРСВБ је имао рекордан број лабораторијских претрага на бруцелозу од преко 350.000 претрага, а утврђена преваленција малих преживара у Републици Српској била је нешто нижа него у Федерацији БиХ и износила је 3,30% наспрам 5,36% респективно. Велики дио домова здравља у Републици Српској тада није имао уведене лабораторијске претраге на бруцелозу, па су се у ВИРСВБ, осим претрага крви животиња, вршиле и претраге узорака крви људи. Ово је био велики допринос ВИРСВБ у сарадњи ветеринарског и јавног здравственог сервиса и огроман допринос у заштити здравља животиња и људи.

During that period, with the active participation of experts from the VRISVB and all veterinary authorities in BiH, an agreement was reached on a joint strategy at the BiH level and an eight-year operational program for the control of brucellosis in BiH (2009-2016) was adopted, which, as a primary measure, provided for the mass vaccination of all small ruminants during the first year of implementation, and the vaccination of breeding stock for the following seven years. The REV-1 vaccine, which contains an attenuated strain, was selected. Brucella melitensis, which, when properly administered, induces lifelong immunity. Enhanced control of cattle for brucellosis is also provided for, whereby, in addition to the mandatory testing of dairy cattle, testing of other categories of cattle over one year of age is also stipulated. This program, already after the first year and mass vaccination of small ruminants, led to a decrease in the incidence of brucellosis in animals and humans. However, the program had its weaknesses, and its implementers were not consistent in its execution, so during the following years, melitococcosis continued to appear, until 2015. 103 cases of human brucellosis and 1,010 small ruminants and 49 cattle were culled in 84 outbreaks in BiH. These data prompted the preparation of a new multi-year program for the control and eradication of small ruminant brucellosis for the period 2017-2021, in the development of which experts from VIRSVB also actively participated. Since mass multi-year vaccination, despite all the identified weaknesses, was the only viable solution, and financial resources for its implementation over the next several years could not be secured, the new program was a continuation of the old one. The main measure remained the vaccination of young ruminants for breeding stock, and some additional activities were added to correct the identified weaknesses. At that time, to control the effectiveness of the vaccination, monitoring of antibody production after vaccination was introduced. After the 2017-2021 program period ended, due to a lack of financial resources for mass vaccination. The continuation of the program with the same main activities has been agreed upon.

VIRSVB is the reference laboratory for brucellosis in the Republika Srpska. For the diagnosis of brucellosis, VIRSVB uses the following testing methods: Rose Bengal, iELISA, cELISA, and the complement fixation reaction. Through two and a half decades of intensive work in brucellosis diagnostics, the number of tests performed at VIRSVB is measured in the millions.

In addition to laboratory diagnostics, which includes monitoring the effectiveness of the vaccination program, the VIRSVB staff conducts epidemiological surveys and assists veterinary organizations, the Ministry of Agriculture's inspection, of Forestry and Water Management of the Republika Srpska in identifying how the infection enters farms, possible routes of its spread, as well as in the detailed determination of brucellosis remediation measures on farms with unclear epizootiological data.

VIRSVB has an established collaboration with the FAO/OIE/WHO Reference Laboratory for Brucellosis (APHA Scientific, United Kingdom) for the diagnosis of brucellosis, with which it participates in proficiency testing.

Салмонелоза

Салмонелоза, заразна болест људи и животиња, са преко 2.600 различитих серотипова Salmonella, is widespread throughout the world. VRSCB has routinely tested samples for salmonella for decades. These are primarily food and animal feed samples, as well as samples from the primary production phase (feces, droppings, litter, surface swabs, bedding, eggs), but also organs from deceased animals.

For Salmonella detection, VIRSVB uses the standard ISO 6579-1 method, and for serotyping, ISO 6579-3. In routine work, five public health-relevant serotypes are serotyped (Salmonella Enteritidis, Salmonella Typhimurium, Salmonela Infantis, Salmonela Virchov and Salmonela Hadar), међутим, ВИРСВБ располаже проширеним панелом антисерума за серотипизацију и других серотипова (Salmonella Montevideo was isolated in 1999 from imported poultry, and Salmonella Kentucky isolated in 2021 in PT testing). The most common Salmonella isolates isolated at VIRSV are: Salmonella EnteritidisSalmonela Infantis Less frequently Salmonella Typhimurium.

Vaccinal strains are occasionally isolated. Enteritidis, Salmonella, which are distinguished by a test for differentiating the wild (field) from the vaccine strain of salmonella.

The results of the work, i.e., the isolates, were confirmed in Slovenia and the Czech Republic, as well as at the Public Institution Institute of Public Health of the Republic of Srpska.

Clostridial infections

Убиквитарне, грам-позитивне штапићасте бактерије из рода Clostridium, cause a wide range of pathological entities in domestic animals. Today, in the genus Clostridium More than 100 species are known, and only about 20 of them are pathogenic. Based on their pathogenicity, they are classified as neurotoxic, histotoxic, enteropathogenic, and Clostridia causing enterotoxemia, as well as atypical Clostridia.

In the territory of the Republika Srpska, histotoxic and enteropathogenic Clostridia, the causative agents of enterotoxemia in large and small ruminants, are most commonly diagnosed.

The role of VIRSVB in controlling Clostridial infections is reflected in the early detection, performance of autopsies, isolation, and identification of the causative agent. Infections with different types Clostridium perfringens In the territory of the Republika Srpska, they most often occur in small ruminants. These include dysentery in lambs and enterotoxemia with degeneration of the kidneys (watery kidney disease, "pulpy kidney disease"). Инфекције овим узрочником забиљежене су спорадично и у узгоју говеда у систему крава-теле.

The diagnosis is made by VIRSVB based on the epidemiological history, clinical signs, pathomorphological findings, and laboratory confirmation based on the isolation of bacteria in pure culture. A suspicion is raised based on the epidemiological history, with information particularly significant regarding age, diet, and season, as well as whether the animals are vaccinated. A justified suspicion can be made based on the peracute course and characteristic pathomorphological changes.

VIRSVB most often receives the carcasses of deceased small ruminants for analysis. In cases of suspected enterotoxemia and water-belly, the carcasses of the deceased animals are in good body condition, and when blood vessels are cut, dark red blood, which is usually uncoagulated, flows out. Hemorrhages and hemorrhagic edema are observed in the subcutaneous tissue. A large amount of serosanguineous fluid is present in the pericardium, pleural cavity, and abdominal cavity. Fibrin clots may be observed in the pericardium. The mucosa of the small intestine, especially the jejunum and ileum, is diffusely reddened, and the lumen most often contains a semi-fluid bloody content. The action of bacterial toxins, which lead to circulatory disturbances, causes a cloudy swelling of the renal parenchyma and an autolytic process that very quickly transforms the kidneys into a red, pulpy mass. This finding is particularly striking and almost pathognomonic for infections with Clostridium perfringens tip D.

VIRSVB isolates and identifies the causative agent from materials obtained during necropsy. By direct microscopy of stained gram-stained smears taken from affected organs and the small intestine, Clostridium perfringens се уочава као велика грам-позитивна штапићаста бактерија, која ријетко формира споре. За изолацију користи се крвни агар, уз стриктне анаеробне услове. Колоније су крупне, глатке, округле, сјајне, окружене карактеристичном зоном двоструке хемолизе. Биохемијска индетификација врши се помоћу API система.

Инфекције хистотоксичним клостридијама на нашем терену јављају се код великих и малих преживара. Најчешће су то шуштавац и парашуштавац. Шуштавац представља акутно или перакутно инфективно обољење углавном млађих говеда, рјеђе оваца, које се карактерише гасним едемом скелетне мускулатуре и често леталним исходом. Узрочник је Clostridium chauvoeiThe material for analysis is most often parts of the affected musculature with internal organs or the carcasses of deceased animals. In cases of cattle death, necropsy is most often performed in the field. The diagnosis is established based on the epidemiological history, clinical signs, pathomorphological findings, and laboratory isolation of the causative agent. The carcasses of deceased animals are distended (postmortem meteorism), and a serosanguineous, frothy fluid drains from the natural orifices. The most prominent pathomorphological changes at necropsy are hemorrhagic infiltrates and edema of the skeletal muscle. The affected muscle has a spongy appearance with gas bubbles. Crepitus is palpated on palpation. The subcutaneous tissue over the affected muscle is firm, infiltrated with hemorrhagic serous exudate, and interspersed with gas bubbles. On the parenchymal organs, grayish degenerative-necrotic and inflammatory foci are noted. Parapneumonia is an infectious noncontagious disease that occurs as a consequence of an infection of the lungs by anaerobic bacteria. The main causative agent of parapneumonia is Clostridium septicum, а појединачно и Clostridium novyi alone or together with Clostridium septicumThe material for necropsy is most often the carcasses of deceased small ruminants, when the history indicates that the possible infection occurred through injuries to the skin and mucous membranes. At necropsy, the pathomorphological changes are very similar to those in the rattlesnake. However, the difference is that in the copperhead, the hemorrhagic component is emphasized, while the emphysematous component is secondary. The localization of the changes is determined by the portal of entry. VIRSVB performs direct microscopy of Gram-stained smears taken from the affected musculature and subcutaneous tissues. Clostridium chauvoei It has the appearance of Gram-negative rod-shaped bacteria with centrally located endospores, with a typical lemon shape. For isolation, liver extract blood agar is used, with incubation under strict anaerobic conditions. On blood agar, colonies Clostridium Shauvoei They are surrounded by a large zone of clear hemolysis. Clostridium septicum Direct microscopy reveals characteristic long, filamentous forms. Endospores are oval and arranged subterminal. On blood agar, colonies have root-like margins and are surrounded by a zone of clear hemolysis.

American foulbrood

American foulbrood is a contagious bacterial disease of the brood of honey bees. Apis mellifera, globally distributed, with significant ecological and economic impacts. There are no drugs that would affect the sporogenic forms of the bacteria, which are the sources of infection of the larvae in their earliest developmental stages. Preventive agents in the form of registered vaccines, serums or genetically modified therapies that can eliminate the causative agent Paenibacillus larvae do not exist. Pharmacological control is not accepted in the same way by the veterinary consensus in many countries. Treatment or prevention (metaphylaxis) with antibiotics is not effective, because it does not affect the spores that are the source of infection for honey bee larvae. Modern beekeeping practices in some countries, with their technological solutions, disregard the natural history of the pathogen and the sustainability mechanisms of bee colonies, thereby increasing the incidence of brood diseases and, consequently, the entire social community.

Early detection of clinically latent forms of infection in infected bee colonies is crucial for establishing control procedures that limit the spread of the infection. Passive monitoring of the clinical forms of American foulbrood in hives has been proven to be insufficiently effective, especially in areas and apiaries with a history of positive findings for Paenibacillus larvae. 
American foulbrood is one of the most prevalent infectious diseases in BiH, considering the number of positive samples submitted to the laboratory. If each sample represents a single hive, with its large population, then the number of individuals participating in complex infectious interactions is enormous. The prevalence of the disease can hardly be determined. The disease has ecological significance, as the spore-forming pathogen remains potentially infectious for many years, and in its reproductive forms, it is very similar to the anthrax pathogen. Bacillus anthracis.

У лабораторијској дијагностици, стандардизацији процедура доказа, комуникацији ризика, едукацији пчелара, учешћима на обукама, конгресима и писању научних и стручних радова у вези са овом незоонозном заразном болести, ВИРСВБ има значајна искуства и ресурсе. Такође, сарадња ВИРСВБ сa Савезом удружења пчелара Републике Српске по овом питању је значајна.

 The implementation of measures against this disease depends on many elements, which fall within the domain of several different stakeholders in this field. The lack of a strategically planned disease control concept, insufficient interest from beekeepers who consider this disease a disgrace, insufficient intensity and quality of veterinary measures, uncertainty in the funding of measures, and outdated regulations, are the reason for the poor indicators regarding the number of positive samples submitted for laboratory analysis.

In the period 2020–2024, VIRSVB tested 4,760 samples for American foulbrood, isolating the causative agent in 29.89% of them.

Q fever

Q fever occupies a significant place in human and veterinary medicine. Since domestic animals represent a significant reservoir Soxiella burnetii, and therefore a potential source of infection for humans, the detection of serological reactors plays an important role in the prevention of this disease in humans. The spread of this zoonosis is favored by the extremely small infectious dose of the causative agent, as well as its high resistance. Q fever represents a major public health challenge, and therefore a challenge for veterinary medicine, in terms of finding an adequate solution for this zoonosis.

Although Q fever in sheep and goats is associated with late abortions and reproductive disorders, such as premature births, stillbirths, and avital offspring, and in cattle was once associated with infertility and metritis, there are generally no specific symptoms related to Q fever. The incidence of Q fever in humans during the winter-summer period is linked to the lambing season. The small infectious dose and high resistance of the causative agent, as well as its persistence in the environment and in vectors, make combating this disease complicated and difficult to implement under our conditions. Due to its ability to cause an epidemic, its environmental resistance, and its mode of aerosol spread, Soxiella burnetii is considered a potential bioterrorism agent. People are predominantly infected during the parturition of animals, especially ruminants, which at that time can excrete a large number of the causative agent. The causative agent is aerosolized and spread by the wind, occasionally traveling long distances, and can thus infect dozens to hundreds of people who are not in direct contact with the animals. Testing for Q fever is mandatory in the event of animal abortions.

The first Q fever epidemic was in 1988 in Omarska (Prijedor), and fourteen years later, a new epidemic occurred in the same epizootiological unit. In addition to the larger epidemics, smaller ones were also recorded in the areas of Gradiška, Laktaši, and Kozarska Dubica, all associated with contacts between humans and small ruminants. In a serological survey in 2001, the VRISVB determined that the seroprevalence in ruminants was 0.5%, and in 2003, it was 0.75%. In the same year, testing of 3,701 sheep in the Banja Luka epizootiological unit revealed the presence of antibodies to Soxiella burnetii in 1.18% of cases. In early 2006, VRISVB surveyed sheep flocks from areas where people had become ill, establishing a seroprevalence of 2.26-29% in those flocks. After nomadic herds passed through the city and its suburbs in 2004, an epidemic occurred with over 300 human cases, predominantly in the Banja Luka area, and to a lesser extent in the Mrkonjić Grad and Čelinac areas.

VIRSVB is the reference laboratory in Republika Srpska and BiH for the diagnosis of Q fever. Testing is performed using the following methods: ELISA test, complement fixation reaction, and Real-Time PCR.

VIRSVB has an established collaboration with the WOAH Q fever reference laboratory (Anses, Sophia Antipolis Laboratory, France) for Q fever diagnostics, with which it participates in proficiency testing.

Diagnosis of transmissible spongiform encephalopathies (TSE) is performed by analyzing brain tissue samples from cattle, sheep, goats, and deer for the presence of the pathogenic prion. 
Diagnosis of bovine spongiform encephalopathy was previously performed using the Prionics Check Western blot immunoblot method and the Prionics PrioStrip immunochromatographic method, while analysis is now performed using an enzyme-linked immunosorbent assay (ELISA). This method meets both domestic and EU regulations governing the diagnosis of transmissible spongiform encephalopathies. The annual monitoring program is based on active and passive surveillance. This program covers cattle over 30 months of age intended for slaughter, as well as cattle over 24 months of age sent for compulsory slaughter or animals that showed symptoms of central nervous system disorders and behavioral abnormalities during antemortem inspections, as well as cattle older than 24 months that died or were euthanized on the premises, during transport, or at the slaughterhouse.
VIRSB also detects the pathogenic prion in the brain tissue of sheep and goats, the causative agent of scrapie (Paraplexia enzootica, Scrapie) and analysis of brain and lymphoid tissue samples from deer for the presence of the pathogenic prion, the cause of chronic wasting disease in deer
(Chronic wasting disease).
VIRSVB is the reference laboratory in the Republika Srpska and BiH for the diagnosis of transmissible spongiform encephalopathies.

Parasites are pathogens that take nutrients from their host and harm its health. The diagnosis of parasitic infections requires laboratory support, as the symptoms of the disease are often non-specific or can be masked by the symptoms of other diseases. In addition to the non-specific clinical picture, many parasitic diseases have a long prepatent period, with a long, completely asymptomatic phase, so it is extremely important for clinicians to be familiar with the epidemiological data on the presence of certain parasites in the areas where the animals have been.

Since its founding, VIRSVB has performed various parasitological examinations, which contribute to elucidating the causes of poor health in animals, as well as in humans. The number of parasitological methods applied at VIRSVB is large, such as fecal examinations, which are of great importance for the detection and control of helminthiases and coccidiosis in large and small ruminants, horses, and pigs, as well as dermatological examinations for the control of mange and lice, and urological and hematological examinations. Of particular importance is the examination of meat for Trichinella larvae using the artificial digestion method, which has contributed to reducing the number of human cases of trichinosis.

VIRSWB is isolating Histomonas spp. and Trichomonas spp., and the diagnosis of parasitic diseases of bees (varroasis, nosemosis, acarosis, etc.) and fish (anisakid larvae). In addition to parasitological methods, VIRSVB also performs molecular tests for the presence of the genomes of certain parasites using the polymerase chain reaction method.

Vector-borne parasitic diseases, such as babesioses and filarioses, have gained importance as a result of global warming and the expanding ranges of the insects that transmit them. In 2015, VIRSVB staff participated in a study on the prevalence of invasive mosquito species in the Vector Net project in BiH, in cooperation with the Faculty of Veterinary Medicine of the University of Sarajevo and the Faculty of Agriculture of the University of Novi Sad. As a result of the project, the presence of Aedes albopictus, and the VIRSVB staff is trained in the morphological determination of mosquito species and genera.

In addition to the above, there are many other parasitic diseases transmitted to animals through trade, as well as through the continuous migrations of wild animals. All of this leads to a complex situation regarding parasitoses, due to the very real possibility of the emergence of parasitic diseases that were once considered exotic to certain parts of the world. The staff of the VIRSVB has always recognized the importance of parasitic diseases in protecting animal and human health and has made an immeasurable contribution to advancing the veterinary profession and the development of science in the field of parasitic diseases. As a result, numerous professional and scientific papers in the field of parasitology have been published.

Trichinellosis

Trichinellosis is a parasitic disease of animals and humans widespread throughout the world which, among animals, is transmitted by consuming meat, and humans become infected by consuming undercooked meat. It is caused by roundworms of the genus Trichinella.

In 1860, during autopsies of humans, James Paget found sand grains in the muscles of the diaphragm. Later, Robert Owen, while microscoping muscle tissue, observed tiny, spiral-shaped larvae of Trichinella embedded within it, which is how he gave the parasite its name. Trichina spiralisWhen the same finding was discovered in pigs, the biological cycle of trichinae was soon discovered, along with the causal link between human infection and the consumption of pork. 
Very quickly, from simple microscopy, the trichinelloscopy or compression method was developed, which involves examining a piece of muscle tissue compressed between two slides for the presence of embedded trichina larvae. This method was immediately introduced as a mandatory measure to protect human health, first in slaughterhouses in Germany in the late 19th century, and then this examination was very quickly adopted as a mandatory inspection for pork in slaughterhouses throughout Europe. The compression method was used for many years, and thanks to it, numerous human epidemics were prevented. However, after human illnesses were recorded from the consumption of meat previously inspected using this method, it was discovered that there was a significant possibility of error and of obtaining a false negative result, due to the small amount of meat examined and the discovery that Trichinella spiralis It is not the only species of Trichinella, but that there are species of Trichinella that are smaller in size (Trichinella brits), or whose larvae do not form a pupa/capsule (Trichinella pseudospiralis) Due to these findings, in the late 20th century, to increase the sensitivity of compression methods, optical search devices were improved and the required sample size for the search was increased. Increasing the number of sections for examination directly affected the length of the examination and the examiner's fatigue. During the 20th century, alternative methods for meat inspection were developed in parallel with the compression method, based on the principle of artificial digestion of the meat. Due to the observed limitations of the compression method (small amount of sample examined, examiner fatigue, low probability of detecting uncapsulated larvae, unsuitability for the large-scale demands of industrial slaughterhouses), In the late 20th and early 21st centuries, the International Commission for Trichinellosis and WOAH-OIE experts stopped recommending it for routine carcass examination, and EU legislation completely restricted this method.

There are several different digestion methods, which are based on the principle of artificially digesting meat samples with HCl and pepsin, thereby releasing larvae from the muscle. These methods allow a large number of pigs to be examined in a relatively short amount of time. The magnetic stirrer digestion method has been designated by the EU as the reference method (EU Regulation 2075/2005, 2015/1375), which is identical to the ISO 18743:2015 method. The digestion method allows for the simultaneous examination of 100-gram samples, shortens the time carcasses are held in slaughterhouses, and reduces the possibility of error due to examiner fatigue. The main advantage of this method is that it allows for, without spending additional time, increase the amount of sample going into the search, which directly and proportionally increases the sensitivity of the trichinae search.

In the Republika Srpska, according to the Regulation on Measures for the Suppression and Eradication of Trichinosis in Animals (Official Gazette of the Republika Srpska 44/10), the artificial digestion method is mandatory. The EU regulation and the regulations of the Republika Srpska and Bosnia and Herzegovina have prescribed one gram of muscle tissue as a sample to be taken for testing using the rapid digestion method on a magnetic stirrer.

The International Commission on Trichinosis has concluded that only a test of a five-gram sample of muscle tissue provides accurate information about the presence of Trichinella larvae in the muscles of the carcass of the examined animal and guarantees the safety of the consumer of the examined meat that he will not be infected with Trichinella, and for endemic areas, which includes the territory of the Republic of Srpska, it is recommended that the test be performed on five grams of muscle tissue. With this in mind, since the introduction of the method of artificial digestion on a magnetic stirrer, the test in the VIRVSVB has been performed on five grams of muscle tissue, which is five times the amount of sample prescribed. Also, the VIRVSVB, in everyday communication, recommends that veterinary doctors in the Republic of Srpska use five grams of sample for the test, especially when the sample they are testing was not taken by themselves. VIRSBV experts, in collaboration with veterinary doctors in the field, determined in 2022 that testing five grams of wild boar meat relatively often reveals low-intensity Trichinella larval infestations, amounting to less than one larva per gram of muscle, which further confirmed VIRSBV's position regarding the amount of sample for examination.

In the period from 1961 to 1964, no case of trichinosis in humans was registered in BiH, and in the period from 1965 to 1991, 1,427 cases of trichinosis were reported. The first cases were identified in 1965 in Banja Luka, when the clinical diagnosis of three hospitalized patients was confirmed in the VIRSVB, by the finding of larvae Trichinella spp. in dried domestic pig meat, using the method of slow artificial digestion using a thermostat. New cases were recorded in 1967, and in 1968 the first large-scale epidemic broke out, again in Banja Luka. Since then, trichinosis has become an endemic disease in BiH, as it began to be regularly diagnosed from year to year. Until 1981, up to 46 sick people were registered per year, and since 1982, trichinosis has become a significant zoonosis with 113 sick people per year. The highest prevalence was in 1982 and amounted to 6.67 cases per 100,000 inhabitants. Human trichinosis in BiH had a constant increase in the period 1965–1991. In the period 1992–2021. In 2008, 2,401 people in BiH fell ill with trichinosis. The average incidence was 80.03 cases, and the average morbidity rate was 2.21% 000. During this period, the morbidity rate of trichinosis in the Republika Srpska and the FBiH was significantly different, when in the Republika Srpska there were 1,789 infected people (average annual 49.38), with an average morbidity rate of 4.48% 000, while in the same period in the FBiH 612 people fell ill with trichinosis (average annual 16.89), with an average annual morbidity rate of 0.87% 000, and the largest number of patients (533) was recorded in 1998. In the period 1992–2001, there were five deaths in Republika Srpska. In the period 2001–2019, human trichinosis has been steadily decreasing, and no new cases were registered in 2020. During 2021, one case was registered, and in 2022, 23 cases were reported due to the consumption of meat products that were not tested for the presence of Trichinella. Until August 2024, there were no new cases of human trichinosis.

Examining the meat of animals for VIRSB, larvae Trichinella spp. were found in muscle samples of domestic pigs, wild boars, bears, badgers, foxes and jackals. During the period 1997-2021, 2,660 trichinosis animal carcasses were reported in the Republika Srpska and 165 in the FBiH, or a total of 2,825 cases in BiH, mostly domestic pigs. During this period to date, the total number of infected animals detected annually has been steadily decreasing, and the largest share of detected trichinosis animals falls on wild boars.

Until 2013, all findings of Trichinella larvae were labeled as findings of larvae. Trichinella spiralis, када су стручњаци ВИРСВБ, у сарадњи с Институтом за примјену нуклеарне енергије Универзитета у Београду (INEP), молекуларном дијагностиком, потврдили епизоотиолошку теорију да на простору Републике Српске и цијеле БиХ, осим T. spiralis, other species of Trichinella are also circulating. Then, samples of Trichinella larvae found in one wild and one domestic pig were molecularly characterized as Trichinella brits and Trichinella pseudospiralis respectively.

Babesiosis

A significant activity of the VIRSVB staff is work on parasitological and molecular typing of the species Babesia ovis. During 2019, an enzootic focus of malignant ovine babesiosis was officially confirmed in the area of ​​the municipality of Rudo. By 2022, epizootiological testing was significantly expanded so that this disease was proven in certain areas of Herzegovina.

Molecular typing and phylogenetic analysis determined the existence of a separate genotype of the species Babesia ovis in the area of ​​the Rudo municipality that differs from the common genotype present in the Middle East. This haplotype is designated as BO53 and is one of 26 proven haplotypes found in the gene database.

The study of malignant babesiosis in sheep was the first in 70 years in Bosnia and Herzegovina. Prior to this study, it was believed that this disease had only historical significance for certain parts of Europe.

When it comes to pathomorphological diagnostics, VIRSVB performs diagnostics of pathomorphological changes in animal tissues. Pathomorphological diagnostics is based on the application of pathoanatomical and pathohistological analysis of organs, or tissues, in order to determine the disease and cause of death of domestic and wild animals. The analysis accepts carcasses and/or organs of animals delivered by the field veterinary service or by the owner.
Veterinary inspectors also send material for analysis, most often due to suspected infectious diseases. Autopsies of carcasses can also be performed in the field, as is the regular case with large ruminants and ungulates. During the processing of the submitted material, samples are taken for microbiological and parasitological analysis. Pathoanatomical analysis is, if necessary, supplemented by pathohistological analysis of tissue.
Complete postmortem diagnostics, in addition to performing an autopsy, includes pathohistological and cytopathological analyses, which aim to make an accurate clinical-pathological diagnosis and precisely determine the cause of death or the presence of a certain disease. The material for pathohistological examination includes material sampled during the autopsy and material sent by the veterinarian or the owner of the animal. Material sent by veterinary doctors or owners
Also, veterinary organizations submit samples of surgically removed tumor tissue for histological analysis. When we talk about the histological diagnosis of tumors in VIRSCV, the most commonly diagnosed are round cell skin tumors (cutaneous lymphoma, plasmacytoma, mastocyte, histiocytoma, melanoma and transmissible venereal tumor), then benign and malignant forms of mammary gland tumors. Also, other neoplasms are diagnosed, i.e. tumors of epithelial and mesenchymal origin.
Pathohistological analyses of organs and tissues can determine specific, or pathognomonic, changes for certain diseases, such as the finding of Bollinger bodies characteristic of smallpox, poultry diphtheria, ecthyma, etc., purulent and non-purulent meningoencephalitis in listeriosis, intracytoplasmic inclusions in neurons in Aujeszky's disease, granulomatous inflammation and the presence of acid-fast pathogens of tuberculosis and paratuberculosis, etc.

VIRSVB prepares daily for microbiological testing
nutrient microbiological media. This is a complex activity that includes various aspects and stages of the process, starting from weighing, dissolving, sterilizing or cooking the nutrient media, and quality control (physicochemical, sterility, productivity, selectivity,
specificity) and handling of reference strains of microorganisms. For the preparation of nutrient media, VIRSVB uses dehydrated media from commercial global manufacturers (over 100 different nutrient media). Approximately 4,800 liters are produced annually.
culture media. For the control of the produced culture media, VIRSCV uses over 40 reference strains of microorganisms, originating from international collections (World Data Centre for Microorganisms WDCM, American Type Cultrue Collection ATCC, Spanish Type Culture Collection CECT, etc.). Annually, VIRSCV produces over 1,000 aliquots of reference microbiological cultures.
In addition to specific, as primary, VIRSVB applies basic methods of isolation and identification of microorganisms in the diagnosis of bacteriological and fungal diseases. This includes isolation and identification on various nutrient media (non-selective, selective, differential) in various temperature and atmospheric conditions of incubation (aerobic, anaerobic, microaerophilic), performing confirmatory biochemical tests (catalase, coagulase, oxidase, biochemical series, etc.) and observation of microscopic preparations (native, Gram, Giemsa, Lefler, etc.). An important aspect of work in a microbiological laboratory is sterilization and
disinfection. Sterilization is defined as the process of complete elimination or destruction of all forms of microorganisms, and disinfection is the destruction or removal of harmful vegetative microorganisms. The main goal of disinfection and sterilization is to reduce the number of microorganisms to such a level that the probability of transmission of infection or contamination is reduced to zero. They are carried out using various physical (heat, ultraviolet light) and chemical processes. In VIRSVB, heat sterilization methods are used, namely dry sterilization for dishes, equipment and utensils (sterilizer at 160°C for one hour), and wet sterilization for nutrient media (autoclave at 121°C for 15/20 minutes or 134°C for 10 minutes). Contaminated material, which includes processed samples, used culture media, glass and plastic dishes, instruments, utensils, etc., is sterilized (decontaminated) at 121°C for 20 minutes. On average, about 270 dry sterilization cycles and 160 wet sterilization cycles are performed annually. Disinfection is performed daily with UV lamps and disinfectants. The effectiveness of disinfection and sterilization procedures is periodically monitored.

During the development of infectious disease diagnostics in the VIRSB, immunological and serological tests have occupied a significant place. In the last three decades, there has been a rapid development of infectious disease diagnostics, with a special emphasis on zoonotic diseases. Over time, new diagnostic procedures have been introduced. For example, slow agglutination methods in the diagnosis of brucellosis have been supplemented over time by newer rapid agglutination methods, as well as immunoenzyme tests, and the complement fixation reaction. Furthermore, the development of new methods resulted in the introduction of the fluorescence polarization test.
The method for diagnosing leptospirosis, the microscopic agglutination reaction (MAT) test, has been used to indirectly detect the presence of leptospirosis infections in animals and humans. The importance of these tests became apparent in 2014, after heavy rainfall and flooding, which created favorable conditions for the maintenance of leptospira in nature.
Serological monitoring in 2014–2015, using the microscopic agglutination test (MAT) with antigens of eight serovars, determined a prevalence of 11.65% in 4,171 ruminant serum samples. Also, determining the peroprevalence in blood samples from dogs and foxes during periods of heavy rainfall provided insight into the importance of carnivores as potential vectors in the chain of spread of eptospirosis. The contribution of VIRSVB in terms of immunological testing is also reflected in the diagnosis of zoonoses, in the testing of human blood serum. At the end of the twentieth and beginning of the twenty-first century, new methods were developed, and, in cooperation with the University Clinical Center of the Republic of Srpska, human blood serum samples were tested, which placed emphasis on the detection of zoonoses in the human population, at a time when this type of diagnosis was not developed in public health. Regarding the diagnosis of Q fever, the introduction of test methods into routine work enabled the detection of this infection in the human population in 2003, when over 300 people fell ill after nomadic herds passed through urban areas. This type of cooperation continued with regard to the diagnosis of brucellosis, leptospirosis, Lyme borreliosis, trichinosis and echinococcosis, until the moment when this diagnosis was established within the framework of public health. The importance of cooperation between veterinary and public health was also reflected in communication and notification in the event of zoonotic infectious diseases. Thus, in 2006, in cases of Q fever in humans, blood samples from animals in their environment were tested, and an eroprevalence of 2.26-29% was determined, which was an indicator of the zoonotic potential of C. burnetii in the environment.

The introduction of new methods and the expansion of the scope of testing also led to the introduction of measures within the framework of legal regulations. In the period 2017–2019, in the submitted serum samples of ruminants with a history of abortion, a prevalence of Chlamydophila obortus of 28.2% was determined, which resulted in the inclusion of this test in the mandatory measures carried out in cases of abortion.
Immunological-serological tests include diagnostics of a wide range of diseases of bacterial and viral etiology, as well as parasitic infestations, regardless of the animal species. Thus, testing for infectious anemia of equines enables diagnostics in
of diseased animals, but also the detection of virus carriers and the reduction of prevalence in the equine population. The introduction of the immunoenzyme test cELISA enabled its use for this purpose, with the obligation to confirm positive results with the AGID test. Immunological tests enabled the diagnosis of CAE disease in imported goats for the first time. This disease continues to pose a threat to goat farming, and immunological tests have an important place in terms of measures and recommendations for combating it. Also significant is the participation in tests for paratuberculosis, BVD and IBR/IPV infections, which are important from the aspect of cattle health protection, as well as for foot-and-mouth disease, which was carried out as part of monitoring.
In the poultry sector, immunological tests have enabled the detection of chlamydial
infection, as well as the determination of specific antibodies in the blood of poultry for diseases such as Gamboro disease, infectious bronchitis and Newcastle disease. Established immunoprophylaxis programs have been optimized based on these tests, with the aim of more adequate protection
flocks and reducing losses caused by infectious diseases. Also, testing for the presence of antibodies against the avian influenza virus has enabled the control of imported flocks.
Changes in climate conditions and modes of transport, including the movement of people and animals, have contributed to the spread of diseases that were previously not present in our area, such as bluetongue. Immunoenzyme testing for West Nile virus indirectly revealed the presence of flavivirus infection. Subsequent testing at the Anses laboratory in France confirmed antibodies to tick-borne encephalitis virus, which proved the presence of this virus in horses in our area for the first time.
Studies within the framework of the training on the development of in-house immunological tests, in collaboration with the Instituto Superiore di Sanità (Rome, Italy), have led to new findings regarding the development of diagnostics and evidence of the presence of antibodies against Trichinella spp. in animal blood serum samples. The development of in-house immunoenzyme and immunofluorescence tests for the diagnosis of leptospirosis, with the aim of serological monitoring, was implemented within the framework of a scientific research project with the Ministry of Science and Technology of the Republic of Srpska. Immunological tests are of great importance in the diagnosis of swine diseases, especially in the serological surveillance of classical and African swine fever. Also important are tests related to Aujeszky's disease, PRRS, circovirus and parvovirus diseases. These tests enable monitoring of the presence of these diseases in pig farming and the introduction of certain control measures on farms.
Regarding bovine brucellosis, continuous serological testing has played a key role in its control for many years, but also in post-vaccination monitoring of small ruminants. These tests have enabled continuous monitoring of a large number of individuals, which has been essential for assessing the effectiveness of vaccination of small ruminants against brucellosis.
In connection with post-vaccination monitoring of rabies in foxes, it has been possible to determine the immune response of vaccinated foxes, which is important in assessing the success of vaccination.

Immunological-serological methods have been among the first to be accredited in the VIRSB since 2011, and the scope of accreditation has expanded over time in line with current diseases. This confirms the importance and role of these tests, which have contributed to the efficiency of disease control and prevention, ensuring timely recognition and response to infectious diseases. This has reduced the risks of their spread and improved animal health management.

In 2009, the VIRSB introduced molecular methods for diagnosing animal diseases into routine work, while it has been conducting rabies testing using the fluorescent antibody technique for decades.
Molecular diagnostic methods include classical PCR and Real Time PCR. Using molecular methods, VIRSB conducts tests on about 25 different animal diseases of viral, bacterial and parasitic etiology, as well as tests on biological materials of animal and other origin. Using molecular diagnostic methods, VIRSB has participated in various monitoring programs, and plays a significant role in the control and suppression of infectious diseases (African swine fever, rabies, avian influenza, brucellosis, bluetongue, classical swine fever, West Nile fever, foot-and-mouth disease). In addition to regular work, VIRSB conducts molecular tests as part of scientific projects, scientific papers and doctoral dissertation experiments. In the field of molecular diagnostics, the VIRSVB is undertaking certain activities related to the procurement of sequencing and bioinformatics equipment, and the introduction of methods for the diagnosis of circovirus, PRRS virus and plague of small ruminants into routine work. In the field of virology and molecular diagnostics, the VIRSVB has established significant cooperation with the World Organization for Animal Health (WOAH, OIE) and EU reference and regional laboratories (Institute of Virology of the Veterinary Faculty in Hanover, Germany; ANSES, France; CSIC INIA, Spain; Zooprophylactic Institute of Padua, Italy; Scientific Institute of Veterinary Science of Serbia, Belgrade, Serbia; Veterinary Specialist Institute "Kraljevo", Serbia; Scientific Institute of Veterinary Science "Novi Sad", Serbia.

In recent years, the staff of the VIRSBV has participated in various projects related to entomological surveys in the territory of the Republic of Srpska. The importance of monitoring the species and density of insect populations, as well as understanding their role in the transmission of vector-borne diseases, is crucial.
Међународни пројекат MediLabSecure у комe је ВИРСВБ учествовао, значајно је помогао
јачању капацитета у овој области, фокусирајући се на:

 – strengthening and harmonizing capacities for preparation and response to health threats that are
mainly associated with vector-borne viruses,
– increasing awareness of the value of integrated surveillance, risk assessment and early warning, in order to prevent and control epidemics/epizootics 
– promoting institutional adaptation for the integrated management of zoonotic diseases of animal origin and stimulating the sustainability of the One Health concept

By participating in the MediLabSecure project, VIRSVB has made significant progress in the area of ​​knowledge acquisition and capacity building related to medical and veterinary entomology. VIRSVB representatives have undergone various trainings, which have resulted in a better understanding of public health, medical entomology and challenges in control, mapping, taxonomy, risk assessment and the functioning of the One Health system. The above elements emphasize the need to integrate human and animal health protection systems. Particularly noteworthy are the very useful MediLabSecure trainings, which include different approaches and can be systematized as work on:

– increasing knowledge and skills in entomology,
– addressing challenges in mosquito control, mapping, taxonomy, risk assessment and elements of monitoring the effectiveness of mosquito control programs 
– integration of the One Health approach to ensure comprehensive health care strategies.

As a result of training and projects, entomological surveys have become a significant factor in the parasitological studies of VIRSB. The most significant activities in these studies include:
– monitoring and determination of the invasive species Aedes albopictus,
– collection and determination of Culicodidae as competent vectors for bluetongue virus
– collection and determination of tick species as competent vectors for bacteria, parasites and viruses.

During 2015, VIRSVB participated in the VectorNet project, together with the Faculty of Veterinary Medicine of the University of Sarajevo and the Agricultural Institute in Novi Sad. The aim of the project was to establish the presence of the tiger mosquito (Aedes albopictus) in Bosnia and Herzegovina. In the Republika Srpska, larvae of this species were established for the first time in Laktaši, and we witnessed that the number and geographical distribution of this species is increasing from year to year. After the bluetongue disease epizootic in 2016, systematic monitoring of Culicoids as competent vectors for this disease began. The largest collection of Culicoids was carried out during the summer of 2017 in the eastern part of the Republika Srpska. The determination of Culicoids proved the presence of the C. obsoletus group and the C. pulicaris complex. Entomological investigations also detected rare species of Culicidae in the Republika Srpska (BiH) that were previously unknown in this area. Entomological investigations include regular monitoring and determination of ticks in the VIRSB laboratories. The largest number of tested ticks was as part of the epidemiological investigation of malignant ovine babesiosis in Podrinje and Herzegovina. In the area of ​​the municipality of Rudo, a field epidemiological investigation in June 2019 established the territorial dominance of the species Rhipicephalus bursa, which was associated with an increased number of clinical cases of ovine babesiosis. DNA of the species Babesia ovis was detected in ticks from the Podrinje and Herzegovina area.

Bovine mastitis is an infectious disease that causes the greatest economic losses in dairy cattle. Due to the importance of mastitis and its role as a public institution, VIRSVB is actively involved in the diagnosis of mastitis. This is reflected in the work of the VIRSVB epizootiology service in the field and the performance of laboratory tests. The epizootiological service of the VIRSVB operates throughout the territory of the Republic of Srpska, which, at the request of the competent veterinary organization, goes out into the field and performs epizootiological investigation and sampling. At both locations where it operates, VIRSVB conducts bacteriological and mycological tests of milk. Bearing in mind that the territory of the epizootiology unit sometimes covers a significant geographical space, the operational activities of the epizootiology service are justified only in case of major damage or when a large number of farms in an epizootiology unit are examined. An additional problem is that, regardless of the availability of veterinary services, some farmers treat dairy cows themselves, and even administer large amounts of antibiotics from the black market, so that veterinarians lose control, not only of mastitis on farms, but also of the amount of antibiotics used on the farm.
Consequently, farmers seek a service when mastitis has already turned into a chronic course and when lactation therapy is almost always unsuccessful.
VIRSVB plays a significant role in the fight against mastitis through the following activities:
- diagnosis of mastitis by methods of pathogen isolation and determination of the number of somatic cells in milk, 
- determination of antimicrobial resistance of the causative agent of mastitis, 
- assessment of hygienic conditions on the farm by methods of swabbing working surfaces and milking equipment on the farm, 
- formation of expert opinion and recommendations with the aim of suppressing mastitis on the farm 
- education of farmers and doctors of veterinary medicine at various gatherings.

Through the activities of regular mastitis diagnostics in VIRSVB, a significant number of bacterial species, fungi and algae were isolated. The most important bacterial causative agents are streptococci enterococci, coagulase-positive staphylococci, coagulase-negative staphylococci, Esherichia coli, Klebsiella spp, Enterobacter spp, Pseudomonas spp, Serratia spp, Tryperella pyogenes, Pasteurella multocida, Corynebacterium spp and nocardia, and fungi include Candida. It is significant that the algae Prototheca was isolated, as well as an atypical, highly slimy strain of the Streptococcus uberis species, which also showed atypicality in the manifestation of biochemical characteristics.

Poultry disease diagnostics is an important segment of VIRSVB's work. It includes work
in the field (epidemiological surveys, clinical examination, sampling) and laboratory
tests:
– патоморфолошка и патохистолошка,
- bacteriological and mycological,
– parasitological, serological and molecular.

In routine work, VIRSVB conducts tests on the following poultry diseases: atypical plague, Gamboro disease, infectious bronchitis, avian influenza, weight loss syndrome, salmonellosis, campylobacteriosis and mycoplasmosis.
As part of serological tests for these diseases, an immunoprophylaxis program is also being developed for all production categories of poultry (mother parent flocks, flocks of consumer layers and broiler flocks). After the primary isolation and identification of the bacterial causative agents, the sensitivity of the isolate to antibiotics is tested. In addition to infectious and parasitic diseases, VIRSVB also diagnoses diseases of non-infectious etiology (technopathies, avitanimosis, etc.). Due to the complexity and sensitivity of poultry production, the diagnosis of poultry diseases requires a multisectoral approach, and in this sense, VIRSVB experts from different fields work on this issue (epidemiology, clinic, laboratory diagnostics, microbiology and animal feed quality).

Beekeeping cannot be compared to other systems of raising animals on farms that produce food for humans. Given that bees are unique social organisms with
complex structure, each individual hive can have a different health status, which
in the final outcome is estimated either as a percentage of survival or annual losses of bee colonies. There are real challenges in understanding basic beekeeping and veterinary practices.
These challenges are often encountered in everyday veterinary practice and are a perennial goal
VIRSVB's activities in this area of ​​health care were focused on the development of diagnostic and
research capacities.
Following the past 30 years (1994-2024), the work of the VIRSVB was focused on diagnostics, development and understanding of the challenges of honey bee sustainability with the aim of creating conditions for better beekeeping in the Republic of Srpska and Bosnia and Herzegovina. Efforts included local and international cooperation, securing grants, as well as improving skills and knowledge in the beekeeping sector. VIRSVB is
established an experimental apiary in 2004, and conducted extensive sampling and
diagnostics, in order to better understand beekeeping practices in the APINET project (2012). The research work of VIRSVB is intensified during the globally recognized golden age of research in beekeeping, at a time when the world learns about unusual and unknown CDC phenomena (Syndrome of Disappearing Bee Colonies 2006) and global losses of bee colonies that have been monitored for years in the Republic of Srpska and Bosnia and Herzegovina in the COLOSS project (2008-2012). VIRSVB used all the laboratory capacities it had, and when it was necessary, it also used the services of other laboratories.
ВИРСВБ је учествовао у бројним пројектима, те се интензивно бавио комуникацијом у медијима и иницирањем преговора о политикама које би требало да допринесу побољшању пракси кроз
примјену стандардизованих и хармонизованих знања.
The networking of VIRSVB with the beekeeping and scientific community was significant, enabling the accreditation of diagnostic methods, verification of results and research into the issue.
sustainability of bees. Toxicological forensic activities were conducted, papers were published, and VIRSVB staff attended conferences (e.g. Apimondia, EurBee) and were in study
visits to prestigious institutes in the USA, Spain, Germany, Russia, South Korea and Argentina. For years, VIRSVB has had a successful collaboration with ANSES, the EU reference laboratory for bee diseases in France.
ВИРСВБ, својом посвећеношћу за унапређење пчеларства, константном едукацијом и
innovations, puts itself in a leading position in the field of diagnostics and sustainable beekeeping in the veterinary sector of the region.
Over the years, VIRSVB staff has been dedicated to improving knowledge and practice in the field of beekeeping through a series of international projects, trainings and cooperation. The professional path of a veterinarian in the field of apidology consists of many elements, in which, as a veterinarian, you need to have professional knowledge, beekeeping experience, the will of a researcher, the activity of a trainer, the skill of a speaker and how much to be a successful advisor. Personal commitment to this part of veterinary medicine in
the work of VIRSVB was reflected in various tasks in the laboratory and in the field, in work with honey bees, but also, if necessary, with wild pollinators. In the laboratory, the expertise includes classic and modern techniques in the diagnosis of bee diseases and parasites, as well as the area of ​​antimicrobial resistance. The spectrum of knowledge includes clinical bacteriology, mycology, virology, protozoa and areas of knowledge of the control of parasites and pests of the bee colony.
The field of toxicological analysis is a special requirement, which is an integral part of the assessment of risks and factors, the levels of which affect the sustainability of the community.
Dealing with challenges in the theory and practice of bee nutrition, selection, health, well-being, vitality, ecological factors of the environment is an integral part of interests within which VIRSVB has support, cooperation and funding sources. Monitoring of new technologies, IoT-related automation, data visualization in GIS, communication with project networks and journalism, have been upgraded over the years in VIRSVB. Dealing with issues of bee nutrition, quality and safety of food for bees, as well as the field of veterinary pharmacology for the needs of the beekeeping sector, are inseparable elements in the process of knowing the sustainability of bee colonies in modern beekeeping, and in the context of public health and the principles of sustainable management and welfare of bees.
In September 2006, there was a study trip by VIRSVB researchers to the USDA Bee Research Laboratory (Beltsville, Maryland, USA), with the aim of improving laboratory and technical field knowledge from the virosphere of bee colonies, financed from the funds of the Unit for the Coordination of Agricultural Projects of the Republic of Srpska, lasting 40 days. Part of knowledge i
the results were used for the preparation of the doctoral dissertation "Detection of virus infections in the Apis mellifera population using molecular biology techniques" (2013).
In October 2008, there was a study trip to the Bee Institute Celle (Germany), focused on the control of bee diseases, with a special emphasis on the American bee brood rot, as part of the BiH Twining project with Germany. This experience significantly contributed to the knowledge and skills in the management and mitigation of bee diseases, especially American bee brood rot, and based on what was learned, VIRSVB launched a multi-year control and prevention activity called "Early diagnosis of American bee rot in apiaries of the Republic of Srpska", supported by the Ministry of Agriculture, Forestry and Water Management of the Republic of Srpska.
VIRSVB was represented at the 2011 OIE Symposium on Bee Disease Diagnosis and Control in Buenos Aires (Argentina).
VIRSVB participated in research projects with the aim of technical cooperation and cooperation in the field of dealing with bees, within the EU project COST action, where it had a MC representative in the COLOSS and SUPERBEE projects.
In April 2015, a representative of the VIRSVB participated in an innovative workshop on the methods of interference nucleic technologies of interfering RNA, called "iRNA and bee health", organized by the Faculty of Engineering and Biological Sciences of the University of Ghent (Belgium), where topics and cutting-edge techniques for understanding and improving the health of bees through RNA interference technologies were discussed.
Also, in March 2015, a representative of VIRSVB participated in a training school with a focus on research called "Methodology for the detection of pathogens in non-Apis pollinators", organized by Martin-Luther-University (Halle-Wittenberg, Germany), which related to the study and the possibility of pathogen exchange between honey bees and other wild pollinators. The acquired knowledge was later used for a national research project in that area in cooperation with the Institute for Genetic Resources of the University of Banja Luka.
VIRSVB participates in the regional project called Biodiversity of the Danube Region
"BEE=Biodiversity 2015 BEE", financed by the Fund for Projects of the Danube Region, in which they participate
collaborators from Montenegro, BiH, Serbia and Austria, with the aim of better harmonizing knowledge and communication with beekeepers, about new scientific knowledge from the sphere of honey bees. A special expansion of practical knowledge about the role of wild pollinators took place at the respectable European Bee Course 2017, held at the Cyprus University of Technology (Limasol,
Cyprus), with the aim of knowing the taxonomy of wild pollinators, under the guidance of world leaders
an expert in that field. VIRSVB's commitment to global challenges in beekeeping and the readiness to, on a very broad scale, approach the place and importance of everything important related to the beekeeping sector, has been strengthened through the regular attendance and participation of the delegated representative of SUPRS in the General Assembly, Apimondia Conference since 2003, including gatherings and announcements in 2005, 2007, 2011, 2013, 2015, in 2019 and 2022, as well as the EurBEE Conference in 2008, 2010, 2012, 2018 and 2022.
In 2019, the VIRSVB representative was the local coordinator of the FAO PSA initiative in beekeeping in Bosnia and Herzegovina.
The volunteering experience of VIRSVB representatives in the beekeeping sector at the international level is also significant, with reference to the work in Phnom Penh, Cambodia (2011), as one of the broader interests for international cooperation and visibility of VIRSVB.
Through joint efforts in the field of research and practice, the book "Good veterinary practice in the apiary" was written (Tomljanović Z., Gajger Tlak I., Santrač V., 2012, Bauer Animal Health), which provides a comprehensive insight into the best practices of veterinary practice in apiculture.
In routine work, VIRSVB conducts laboratory tests on the following pathogens and diseases of bees: American bee brood plague, European bee plague, nosemosis, varroosis, acarosis, calcareous brood, stone brood, Aethina tumida, Tropilaelaps spp., virus of deformed bee wings, black queen virus, mixed brood virus, acute bee paralysis virus and chronic bee paralysis virus. 

VIRSVB performs analyses of biochemical and hormonal parameters from blood serum and plasma, for the purposes of assessing the health and metabolic status of domestic animals. These analyses are also performed within the framework of scientific projects, monitoring and experiments for the needs of postgraduate studies.
studies
За биохемијске анализе користи се аутоматски биохемијски анализатор (A25 BioSystems),
intended for in vitro diagnostics, specially designed for clinical biochemistry. For this analyzer, the samples for analysis are blood serum, urine, plasma and cerebrospinal fluid.
The immunochemical analyzer (TOSOH AIA 360) is used to determine hormone levels in blood serum and plasma.
Laboratory examination of blood and other body fluids occupies a very important place in
diagnostics of organic diseases of all types of domestic animals. Biochemical indicators are indicators of nutritional status and metabolic balance in the organism, during different periods of the production-reproductive cycle. Increased or decreased values ​​of a certain parameter indicate the level of damage to the cells of certain tissues, which further indicates a reduced
functional capacity of the organ.
The metabolic profile of cows is the determination of biochemical blood parameters important for assessing health status and production performance in high-milk cows.
cow. The special importance of metabolic profile analysis is reflected in the assessment of the usability
nutrients in cow diets, in different phases of the production and reproductive cycle,
i.e. their energy, protein and mineral status. By examining the metabolic profile, it is possible to detect disorders in the functioning of, for example, the liver, and thus timely
implement appropriate veterinary-medical measures. Biochemical parameters for assessment
metabolic profile in cows are: glucose, betahydroxybutyric acid, total protein
albumin, urea, calcium, phosphorus, magnesium, cholesterol, triglycerides, bilirubin,
triglycerides, AST and gGT. For the assessment of energy metabolism, biochemical parameters are glucose
and betahydroxybutyric acid, for the assessment of liver and renal function proteins, albumin and urea, for the mineral status Ca, Mg and P, for the assessment of the functional state of the liver bilirubin, ALT, AST
and gGT, and amylase for liver and pancreas status..
Laboratory analysis of hormone levels in the blood assesses metabolic and
health status in animals, as well as their impact on reproductive performance in
breeding animals.
У оквиру ових испитивања, ВИРСВБ је учествовао у научно-истраживачком пројекту испитивања нивоа хормона у млијечном и крвном серуму крава, као и испитивању биохемијских
parameters in high-milk cows that had elevated levels of aflatoxin M1 in milk.

Nearly two-thirds of human diseases originate from domestic and wild animals, known as zoonoses. Many diseases pose a risk to public health worldwide, making it imperative to control these diseases at local and global levels.
The One Health concept is based on the awareness of the need to protect public health, acting primarily on the prevention and control of pathogens within human and animal populations in the environment in which they live. It is a collaborative, multisectoral and transdisciplinary approach, working at local, regional, national and global levels, with the aim of achieving optimal health outcomes, recognizing the interconnectedness between people, animals, plants and their shared environment. The main areas of work in which the concept is
"One Health" is particularly relevant to:
– control of zoonoses,
– food safety,
– combating antibiotic resistance, 
– environmental protection and – animal welfare.

The “One Health” concept began to be implemented after the signing of a joint document between the World Health Organization (WHO), the World Organization for Animal Health (WOAH) and the Food and Agriculture Organization (FAO). Since 2022, the United Nations Environment Agency (UNEP) has joined the initiative through a memorandum of understanding. This document outlines mutual responsibilities and goals in this area. The most effective and cost-effective solution for protecting people is to combat all zoonotic pathogens in animal populations. This requires a strategic approach, organization, management and control with adequate financial investment and engagement of available resources.
human and material resources from the public and private sectors. This implies establishing new mechanisms for mutual information and synchronized joint action.
The "One Health" concept is partly covered by the following regulation:
– Regulation on the method of mutual notification of health and veterinary services (Official Gazette of the Republika Srpska 46/13),
– Rulebook on the method of monitoring zoonoses and zoonotic agents (Official Gazette of BiH 46/10, 96/13).

VIRSVB is a signatory to the protocol on cooperation regarding mutual information on data in the field of zoonoses control with the Public Health Institution Institute for Public Health of the Republika Srpska (PHI IJZRS). This protocol defines the rights and obligations of the signatories regarding the exchange of data related to diseases, conditions, epidemics and deaths resulting from the occurrence of zoonoses, i.e. anthropozoonoses and their causative agents, as well as the monitoring of diseases and their causative agents, monitoring the resistance of zoonoses to antibiotics, epidemiological and epizootiological studies of the methods and causes of the occurrence of zoonoses that are transmitted through contact with a sick person, sick animal or food. As a result of these activities, there is regular mutual information on the occurrence of zoonoses. Every month, based on data collected from its own laboratories and field data, the VIRVS compiles a Bulletin on Zoonoses, where certain zoonoses are registered, which, as information, is important for all bodies and institutions within veterinary medicine, but also for health institutions that can take appropriate measures based on this official information. In addition to zoonotic diseases diagnosed in animals, the Bulletin on Zoonoses also publishes data on the detection of zoonotic pathogens in food and water. Public health is particularly highlighted in the "One Health" concept, because numerous pathogens are transmitted through food and water. The most significant zoonoses that are occasionally registered in the Republika Srpska are: brucellosis, salmonellosis, Q fever, leptospirosis, trichinosis, etc. A monthly report on all notifiable infectious animal diseases is published by the Veterinary Department of the Ministry of Agriculture, Forestry and Water Management of the Republika Srpska. This report publishes data on infectious animal diseases of all types, both zoonotic and other infectious diseases. The Public Health Institute of the Republic of Srpska publishes monthly reports on the incidence and death of zoonoses, which are regularly submitted by the Veterinary Veterinary Service. Veterinary Veterinary Service
exchanges reports on zoonoses with the Public Health Institution, Health Center Banja Luka.
VIRSVB promotes education and awareness about the "One Health" concept and its importance in addressing health challenges, including zoonoses, antimicrobial resistance, food safety and environmental protection, through numerous educational events and public events.
media appearances. VIRSVB actively develops and proposes policies and regulations that support the "One Health" concept and promotes integrated approaches to addressing health challenges and environmental protection.
Antimicrobial resistance is a global problem in medical and veterinary microbiology. It has been studied for decades and is considered to be one of the greatest challenges in the near future for medicine.
The VIRSB regularly determines antimicrobial resistance to isolated pathogens. In routine work, many isolates showing multiple resistance to various antimicrobial agents were detected. One of the significant discoveries was the discovery of methicillin-resistant strains of Staphylococcus aureus from cow's milk. An increase in highly resistant strains of Escherichia coli was observed in cow's milk and intestinal contents of calves with neonatal diarrhea. No less significant problem are the findings of resistant strains of Staphylococcus pseudintermedius in dogs with clinically manifest pyoderma.

A significant number of laboratory tests in the field of food safety at VIRSVB relate to microbiological tests, which include testing samples of food, animal feed, water, and microbiological purity.

When it comes to microbiological testing, VIRSVB annually tests, on average, around 20,000 samples, including 13,000 food samples (65%), 500 animal feed samples (2.5%), 500 water samples (2.5%) and 6,000 microbiological purity samples (30%).

The users of the services of the above microbiological tests are food business entities, inspection (veterinary, food and border) and veterinary organizations. Within the self-control system, food business entities in most cases have a concluded cooperation agreement with VIRSVB, on the basis of which regular sampling and microbiological testing of samples is carried out. Of the total number of samples, about 95% are from self-control. Another type of sample delivery is official control by the inspection, which accounts for about 3-4% of analyzed samples, and 1-2% of samples are samples from monitoring and delivered by veterinary organizations.

The legal regulations that define this area are as follows: 
– Food Law (Official Gazette of the Republika Srpska 19/17),
– Law on Veterinary Medicine in the Republika Srpska (Official Gazette of the Republika Srpska 75/17, 63/24),
– Law on the Protection of the Population from Infectious Diseases (Official Gazette of the Republika Srpska 90/17, 42/20, 98/20, 63/22),

Food Law (Official Gazette of BiH 50/04).

The VIRSB analyzes the obtained results of microbiological tests, compares them over many years, both with its own and with the results of other researchers, publishes them, and uses them as a basis for making proposals, guidelines and strategies in the field of microbiological testing of food, animal feed, water and microbiological purity. Also, as an active participant, the VIRSB makes a significant contribution to scientific and professional discussions and meetings, and when considering and adopting regulations at the level of the Republic of Srpska and Bosnia and Herzegovina, which relate to this area.

Water

VIRSBV performs microbiological testing of water samples, which represent drinking water originating from animal farms (mostly broiler chicken and dairy cow farms) and food production plants (mainly meat, fish and milk industries). These samples originate from water pipes and wells. The role of VIRSBV from the aspect of microbiological testing of water is, on the one hand, ensuring the health of animals by supplying them with safe water, and on the other hand, ensuring human health by controlling water as a raw material in the food production process. On average, about 70% of satisfactory and about 30% unsatisfactory water samples are determined annually. The regulation according to which VIRSBV tests water samples is as follows:
– Regulation on the health safety of water intended for human consumption (Official Gazette of the Republika Srpska 88/17, 97/18),
– Regulation on the sanitary quality of drinking water (Official Gazette of BiH 40/10, 43/10, 30/12, 62/17). 

Микробиолошка чистоћа

Microbiological cleanliness samples include samples from the working and production environment in food production and trade, and these are swabs of equipment, devices, utensils, work surfaces, work clothes and hands of workers in production and trade in facilities and means of transport that come into contact with food. A significant number of samples are used for this segment of microbiological testing, almost one third of which VIRSVB pays special attention to. On average, about 95% of satisfactory and about 5% of unsatisfactory microbiological cleanliness samples are determined annually. The cause of unsatisfactory samples is in the absolute majority an increased number of microorganisms, significantly less an increased number Enterobacteriaceae, док патогени микроорганизми Salmonella and Listeria have not been determined. The regulation according to which VIRSVB tests samples for microbiological purity is as follows:
– Rulebook on the procedure for determining microbiological purity criteria (Official Gazette of the Republika Srpska 74/18),
– Regulation on microbiological criteria for food of animal origin (Official Gazette of the Republika Srpska 69/19, 102/19, 30/22),
– Guidelines for the interpretation and implementation of the Rulebook on Microbiological Criteria for Food (Food Safety Agency of BiH, 2023).

Animal food

Animal feed samples that are microbiologically tested in the VIRSBV are represented in a very small number compared to other sample categories. They are mainly feed mixtures and feeds of plant origin, and to a lesser extent of animal origin, pelleted animal feed, premixes and vitamin-mineral supplements. On average, about 70% of satisfactory and about 30% unsatisfactory animal feed samples are determined annually. The cause of unsatisfactory samples is in the absolute majority an increased number of microorganisms and an increased number of yeasts and molds, while pathogenic microorganisms are the reason in 0.5% of cases. The regulation according to which the VIRSBV tests animal feed samples is as follows:
– Rulebook on microbiological criteria in animal feed (Official Gazette of BiH 67/12).

Food

Food samples represent the largest number of samples submitted for microbiological testing and come from entities in the food business at the industrial and wholesale level (dairies, slaughterhouses, retail chains, etc.), but also from entities in the food business at the artisanal production and retail level (butcher shops, family farms and agricultural producers, restaurants, bakeries, fast food establishments, etc.).
The structure of the analyzed samples is very diverse, which includes the following categories:
– meat and meat products,
fish and fish and seafood products,
– milk and milk products,
– cereals, confectionery, mill and bakery products and pasta,
– honey and honey products,
– eggs and egg products,
– semi-finished and ready-made meals,
– fruits, vegetables and fruit and vegetable products, 
– soup concentrate,
– tea and coffee, 
– soft drinks, beer and ice, 
– адитиве, ароме, боје и зачине,
– baby food and edible fats and products.

Microbiological testing of food is carried out using BAS ISO reference methods, introduced into routine work in 2012, in accordance with Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs.

When it comes to food samples, the most common tests are on three microorganisms, approximately 6,000 (46%) each. Salmonella sppand Listeria monocytogenes and about 2,500 (19%) on Escherichia coli.

On average, about 97% of food samples are found to be satisfactory and about 3% to be unsatisfactory annually. The regulations according to which the VIRSB conducts food sample testing are as follows:
– Regulation on microbiological criteria for food of animal origin (Official Gazette of the Republika Srpska 69/19, 102/19, 30/22),
– Regulation on microbiological criteria for food (Official Gazette of the Republika Srpska 109/12),
– Rulebook on Microbiological Criteria for Food (Official Gazette of BiH 11/13, 79/16, 64/18),

– Guidelines for the interpretation and implementation of the Rulebook on Microbiological Criteria for Food (Food Safety Agency of BiH, 2023).

According to the current regulations, the determination of histamine is classified as a microbiological test. Histamine is a biochemical product found in certain fishery products, and its presence may pose a health risk if consumed in large quantities. Quality control of these products is of utmost importance, both at border crossings and in domestic trade. Detection and quantification of histamine in fish and fish products is performed by HPLC method (reference BAS ISO method). The VIRSVB performs 100-150 analyses annually to determine the amount of histamine.

The system of ensuring and controlling the quality of food and feed in the world today is based on an integrated approach that monitors their production at all stages, from production to the consumer. The central European organization for risk assessment and ensuring the safety of food and feed is the European Food Safety Authority (EFSA). The rapid flow of information is ensured by the Rapid Alert System for Food and FEED (RASFF). The work of these organizations and the successful assurance and control of the quality of food and feed would be unthinkable without a network of authorized and/or accredited laboratories, which play a key role in implementing food and feed safety control and protecting public health, and help national authorities determine priorities in food safety control and direct programs accordingly.
In 2000, VIRSBV initiated the development and introduction into routine work of physicochemical methods for determining the quality parameters of food and animal feed. Today, with a total of 80 physicochemical methods, VIRSBV invests significant resources in the further development and introduction of new methods and analysis techniques, achieving continuous progress and satisfaction of service users.
In addition to the usual laboratory equipment used in methods based on basic analytical techniques, highly sophisticated equipment is also involved in the quality control of food and animal feed. This equipment not only supports existing methods, but also enables the development and application of new methods that require high precision, sensitivity and testing capacity. The basic parameters of food and animal feed quality most often determined are water content, ash, cellulose, protein, free and total fat, density, milk fat, salt, phosphorus, calcium, acid value and acidity, peroxide value, HMF, diastase, water-insoluble matter, free acid content, pH, reducing sugars and sucrose, electrical conductivity, equantometric degree, freezing point, food additives (nitrites, nitrates, phosphates, sorbates and benzoates).
The determination of the quality and healthiness of food and animal feed is carried out according to the regulations and standards in force in the Republika Srpska and/or BiH. The Ministry of Agriculture, Forestry and Water Management of the Republika Srpska and the Food Safety Agency of BiH are the central authorities in this area, which adopt regulations covering analysis methods, quality standards and quality parameters of food and animal feed. A major step in harmonization with the European Union in the area of ​​ensuring and controlling the quality of food and animal feed
animals is represented by the Food Law, adopted in 2004 at the level of BiH (Official Gazette of BiH 50/04), and in 2017 at the level of Republika Srpska (Official Gazette of Republika Srpska 19/17). This law regulates the general conditions for the safety of food and feed, the obligations and responsibilities of entities in the food business, including traditional products, as well as other issues significant for the safety of food and feed, in order to protect the life and health of people, the environment, consumers and the efficient functioning of the market. In the field of food quality, the following rulebooks are most often used:
– Regulation on minced meat, semi-finished products and meat products (Official Gazette of the Republika Srpska 46/15),
– Rulebook on minced meat, semi-finished products and meat products (Official Gazette of BiH 82/13, 84/17),
– Regulation on Market Standards for Poultry Meat (Official Gazette of BiH 74/14),
– Regulation on Food Additives (Official Gazette of BiH 33/18, 6/21),
– Regulation on Food Additives (Official Gazette of the Republika Srpska 96/20),
– Regulation on the Provision of Food Information to Consumers (Official Gazette of the Republika Srpska 9/18),
– Regulation on the Provision of Food Information to Consumers (Official Gazette of BiH 68/13),
– Regulation on Lubricating Greases (Official Gazette of BiH 21/11),
– Regulation on milk products and starter cultures (Official Gazette of the Republic of Srpska, 19/24) and Regulation on milk products and starter cultures (Official Gazette of BiH, 21/11, 25/12, 17/19),
– Regulation on condensed milk and powdered milk (Official Gazette of BiH 21/11),
– Regulation on the quality of fresh raw milk and conditions for the work of authorized laboratories (Official Gazette of the Republika Srpska 81/15, 46/18).
– The Regulation on the Provision of Food Information to Consumers (Official Gazette of the Republika Srpska 9/18 and Official Gazette of BiH 68/13) is of vital importance because it enables the identification of potential hazards and irregularities in the food chain, thereby protecting human health and preventing major economic damage.

By implementing preventive measures, such as stricter monitoring and rapid response to potential risks, it is possible to maintain the integrity of the food supply chain and ensure that food is safe and of good quality for consumers. This systematic control, which includes regular analysis of food and feed samples, helps to detect products of questionable and unsatisfactory quality, as well as the presence of substances that may, in larger quantities, endanger human or animal health. This improves the overall quality of food and the reliability of food on the market. Thanks to advanced technology, analytical capabilities and capacity, various chemical substances are detected and quantified with high precision and reproducibility, which is necessary for complex tests in the field of food and feed safety. Tests for the determination of additives are highlighted as particularly relevant and important. Food additives are substances added to food to improve its organoleptic, nutritional or technological characteristics. They are used to improve the colour, flavour, texture or shelf life of food. Additives include colours, preservatives, antioxidants, emulsifiers, stabilisers, sweeteners and flavour enhancers. The control of additives is essential, as some of them can be harmful to human health if used in large quantities or if not applied appropriately. Therefore, regular control of food additives is mandatory to ensure that they are used within permitted limits and do not endanger the health of consumers.
For the determination of additives, various analytical techniques are used to determine their presence, concentration and possible interaction with other food ingredients. These include techniques such as HPLC (high performance liquid chromatography), ELISA (enzyme-linked immunosorbent assay), spectroscopy, as well as chemical methods of analysis. This approach allows for obtaining precise and reliable results, which are essential for the protection of public health and compliance with the Regulation on Food Additives (Official Gazette of the Republic of Srpska 96/20) and the Regulation on
food additives (Official Gazette of BiH 33/18, 6/21). The possibilities for determining additives are great and work is constantly being done to introduce new technologies in order to improve the control of food additives on the market. VIRSVB tests nitrites (E 249-250), nitrates (E 251-252), phosphates (E 338-452) and benzoic and sorbic acids (E 200-219), and methods for the analysis of sulfur dioxide and sulfites (E 220-228), as well as for the analysis of colors in food, are being developed.

Raw milk quality involves assessing the quality through chemical composition (milk fat, crude protein, lactose, dry matter without fat) and hygienic correctness (somatic cell count and total bacterial count). In a large number of countries, milk buyers exclusively use flow cytometry, i.e. IR spectrophotometry for raw milk analysis. Wanting to follow this trend, enable improvement of raw milk quality and increase its production, VIRSVB, in 2004, with the assistance of the Ministry of Agriculture, Forestry and Water Management of the Republika Srpska (MPSW RS), established a laboratory for testing raw milk. Initially, the instruments for raw milk analysis were located at two locations, Banja Luka and Bijeljina, but, due to the idea of ​​starting a quality control system, all instruments were placed at the Banja Luka location.
The methods used for raw milk analysis are as follows:
– determination of the total number of bacteria by flow cytometry (according to the instructions of the equipment manufacturer FOSS, Denmark) on the highly sensitive Bactoscan instrument,
– determination of the chemical composition of milk by IR-spectrophotometry (according to the instructions of the equipment FOSS, Denmark and according to ISO 9622:2015 Milk and liquid milk products – Guidelines for the application of mid-infrared spectrometry) on the Milkoscan instrument and determination of the number of somatic cells by flow cytometry (according to the instructions of the equipment manufacturer FOSS Denmark and according to BAS EN ISO 13366-2/Cor1:2008 Milk
– Somatic cell counting – Part 2: Guide for the operation of fluoro-opto-electronic counters (EN ISO 13366-2:2006/AC:2007, IDT) on the Fossomatic instrument.

Raw milk testing according to the Regulation on the quality of fresh raw milk and the conditions for the work of an authorized laboratory (Official Gazette of the Republic of Srpska 81/15) is carried out on two samples per month for each producer, determining the chemical composition and total number of bacteria, and the number of somatic cells once a month. The decision of the Ministry of Agriculture, Forestry and Fisheries of the Republic of Srpska sets the adequacy limits for the percentage of milk fat and protein that eliminate milk samples, which in that case are not used in the calculation for the class. The staff responsible for milk analysis is trained by an authorized equipment repairer, who also installed the analysis instruments and has been responsible for maintaining the equipment for many years. Any change in the software of the instruments themselves requires installation by
service technician and in-service testing, as well as staff training for new software versions. Part of the staff is trained to monitor the quality control of the Bactoscan FT 100 and 150H and CombiFoss 200H, which enable the processing of a large number of samples in a short period of time, and the laboratory also has the accompanying equipment necessary for the preparation of samples for analysis and their storage.
VIRSVB, in cooperation with IT experts, has developed the application software "Mlijekosoft", which is used to collect and process data obtained from analyses on the instruments themselves.
Using barcode labels, where BA farm numbers are used to protect the identity of the owner of the milk samples, it is reviewed impartially, and the owner's identity can only be seen after the analyses are completed and the values ​​are downloaded into the application software.
Before the system was launched in 2009, more than 700 people were trained to take samples of raw milk in the Republika Srpska. In addition to the people nominated by dairy farmers/milk purchase organizers, training was also provided to employees of the Ministry of Forestry and Fisheries of the Republic of Srpska (the department for providing professional assistance), who later (2018) took over most of the sampling work. The launch of the milk control system led to an intensification of milk analyses, so that in 2008, 61,165 raw milk analyses were performed (at the request of dairy farmers/purchase organizers and individuals), and the following year, when the system was launched, 162,012 analyses were performed. In 2010, the methods used in the milk quality testing laboratory were first accredited by the Accreditation Institute of Bosnia and Herzegovina (BATA), and have been in accreditation status since then. In the period 2009–2023, 1,723,000 analyses of raw milk samples were performed in VIRSVB, and since the very beginning, around 2,500,000 analyses. The largest number of raw milk suppliers was recorded in March 2009, when there were around 9,500 suppliers. This number has been decreasing in the following period, which is a tendency that is noticeable in all neighboring countries that have a similar pattern of raw milk control. At the end of 2023 and the beginning of 2024, the number of suppliers in the quality control system was around 3,200.
Since 2004, and before obtaining accreditation status, the quality of analyses, i.e. the entire work process, has been controlled. This refers to the recommended daily, weekly, monthly and periodic controls, both by the manufacturers of measuring equipment, and controls recommended by ISO standards. In order to confirm the work of the laboratory, comparisons were made with other laboratories within the framework of interlaboratory comparisons and PT schemes, organized by the Reference Laboratories from Croatia (Zagreb) and Slovenia (Ljubljana). Also, the control of instruments was carried out with secondary reference material, prepared in the Reference Laboratory for Milk of the Faculty of Agronomy in Zagreb. VIRSVB has successfully passed several FVO inspections related to the export of milk and milk products, the first of which was in 2014, without any complaints about the work of the laboratory.
The milk control system has in many cases been recognized and accepted by milk producers as a mechanism for improving milk production and quality, so that in January 2014, there were 52.30% of producers who achieved standard quality milk, and
In 2024, there were 89.50% of producers who had milk of standard quality. This number refers to producers who have milk of standard quality (geometric mean of the number of somatic cells of three-month tests and geometric mean of the total number of bacteria of two-month tests), and not to the total amount of milk produced in liters. Of course, these values ​​​​variegate during each year, so that quality declines were observed mainly in the summer period, which is unfavorable due to high temperatures because they also affect production animals, but also make it difficult to manipulate milk after milking (cooling and transport). Also, the winter period is unfavorable in some cases due to extremely low temperatures.

Sensory (organoleptic) food analysis is the process of evaluating the organoleptic properties of food using the human senses, such as appearance, color, odor, taste, consistency, and texture. This type of analysis is of utmost importance, as it provides information about the quality and acceptability of food for consumption. Irregularities in the sensory properties of food may indicate the presence of deficiencies in production, compliance with quality standards, or potential health risks.
If food does not meet the requirements in terms of sensory properties, this can have significant consequences for human health. For example, the presence of an unpleasant odor or taste can indicate possible contamination or inappropriate storage conditions, which can lead to poisoning and/or other health problems. Therefore, regular and adequate sensory analysis is crucial for the safety and quality of food that reaches consumers.
The VIRSB conducts approximately 300 sensory analyses of food annually. Laboratories play an important role in the assessment of food declarations by providing an opinion on their completeness and accuracy. Declarations that are incomplete or contain errors can lead to
mislead consumers, regardless of whether the errors were intentional or accidental. Food business operators who place packaged food on the market are obliged to provide precise and clear information on the declaration, as they bear all responsibility when it comes to informing consumers.
The opinion on food declaration is of utmost importance because it allows food business entities to identify and timely eliminate deficiencies and errors in accordance with the requirements of the Regulation on the Provision of Food Information to Consumers (Official Gazette of the Republika Srpska 9/18 and Official Gazette of BiH 68/13). Timely and continuous control of food declarations significantly contributes to consumer trust in the information provided on the declaration and the protection of consumer health. On an annual basis, the VIRSB issues about 15 opinions on declarations.

Every product placed on the market must comply with food safety standards set out in regulations. The standards refer to legally defined amounts of substances that may be harmful to the health and well-being of consumers. The growing role of chemical
The use of chemicals in agricultural and other food production and processing has made it necessary to monitor residues and contaminants in food and animal feed. The most important contaminants in the food industry are residues of antibiotics and chemotherapeutics, pesticides, polychlorinated biphenyls, dioxins, mycotoxins, polycyclic aromatic hydrocarbons (acrylamide, melamine), heavy metals and toxins of plant origin.

ВИРСВБ у овој области врши сљедеће активности:

Veterinary medicines

The issue of monitoring the presence of residues of veterinary drugs is complex and demanding given the large number of groups of substances (due to the difference in the chemical composition of individual compounds) and the significant impact on metabolic changes that the presence of residues has on the final product.
To determine the presence of residues of veterinary drugs, VIRSVB applies orientational microbiological (Premi test and 6-plate test), biochemical (Enzyme-Linked Immunosorbent Assay ELISA) and instrumental methods (high-performance liquid chromatography with mass detection UPLC-MS/MS).
VIRSVB annually analyzes approximately 1,500 samples of meat, offal, milk and eggs for the presence of residues of veterinary drugs, namely analyses for the presence of tetracyclines, sulfonamides, penicillins, cephalosporins, aminoglycosides, quinolones and macrolides (qualitative analyses).
The screening ELISA method is used for the analysis of sulfonamides and chloramphenicol in milk, eggs and urine, florfenicol in milk, and the high-performance liquid chromatography with mass detection (UPLC-MS/MS) method is used for the analysis of tetracycline in milk.
The laboratory develops and evaluates new analytical methods that enable a wider range and spectrum of determination of chemical compounds in various foods, following increasingly strict rules that constantly reduce the levels of maximum permitted amounts of residues in various types of food, which are defined by the Regulation on maximum permitted amounts of pharmacologically active substances in products of animal origin.
(Official Gazette of BiH 84/22, 33/24).

Mycotoxins
Mycotoxins are toxic substances produced by various types of molds, mainly Aspergillus, Penicillium and Fusarium. Factors that influence the production of mycotoxins are: fungal strain, type of substrate, ambient temperature, humidity and water content in the product, as well as its degree of maturity. In addition to health risks and diseases, mycotoxins lead to huge economic losses. VIRSVB tests 3,000-4,000 samples annually for the presence of mycotoxins, of which 60% are for aflatoxin M1 and B1, and the rest for ochratoxin, T-2 toxin, deoxynivalenol, fumonisin and zearalenone. This area is regulated by the Regulation on maximum permitted quantities for
одређене контаминанте у храни (Службени гласник Републике Српске 32/21 и СГ БиХ 68/14, 79/16,
9/17, 84/18), Regulation on undesirable substances in animal feed (Official Gazette
BiH 72/11, 70/16) and the Guidelines on the presence of mycotoxins pheoxyvalenol, zearalenone,
охратоксина А, фумонизина Б1+Б2 и токсина Т-2 и ХТ-2 у производима намијењеним за исхрану животиња (Агенција за храну БиХ, 2015). Присуство афлатоксина М1 у млијеку, поред оријентационе методе, од 2014. године анализира се и потврдном методом течне хроматографије високих перформанси с масеном детекцијом, UPLC-MS/MS.

Pesticides
The use of pesticides in agriculture has led to a significant increase in food production. The most important pesticides are insecticides (agents for destroying insects, parasites, flies, ticks, etc.), rodenticides (destruction of rodents), herbicides (agents for protecting against weeds), fungicides, etc. Most pesticides are toxic to domestic animals, fish, bees and humans, and some of them are carcinogenic due to prolonged consumption. The source of exposure for animals can be their direct treatment with pesticides, inhalation of polluted air and ingestion of contaminated feed, with all of the aforementioned routes of intake resulting in intoxication of animals and accumulation of these compounds in food products of animal origin. Chronic exposure to low doses of pesticides in humans results in possible toxic effects, the most pronounced of which are neurotoxic, immunotoxic, and
and genotoxic effects, and toxic effects on the reproductive system.
The groups of pesticides that are most extensively and most researched in the context of contamination of meat and meat products are persistent environmental pollutants, with the following five subgroups of these compounds most commonly analyzed: organochlorine pesticides (OCPs), organophosphorus pesticides (OPPs), synthetic pyrethroids (PYRs), carbamates, and triazines. Examples of organochlorine insecticides are: DDT- dichlorodiphenyltrichloroethane, dieldrin, aldrin, chlordane, lindane (γ isomer of hexachlorocyclohexane), endosulfan, etc.
The provisions of the Stockholm Convention from the 1980s banned the use of many compounds from this group (DDT, dieldrin, chlordane) in agricultural purposes, as well as in households, after it was established that they are extremely stable and lipophilic, and that they accumulate in animal fat. These compounds are considered endocrine disruptors and carcinogenic chemicals. In order to protect consumers from exposure to unacceptably high levels of pesticide residues in food and animal feed, regulations have set maximum permitted levels for different types of food, in most cases expressed as the sum of the concentrations of the main compound and
its metabolites. Due to the above, monitoring the presence of pesticides becomes mandatory, both in foods of plant and animal origin, as well as in animal feed, which is regulated by the Regulation on maximum levels of pesticide residues in food and feed and on food and feed (Official Gazette of the Republic of Srpska 55/20, 59/22) and the Regulation on maximum levels of pesticide residues in
levels of pesticide residues in and on food and feed of plant and animal origin
(Official Gazette of BiH 48/21).
Since 2018, the VIRSB has been conducting testing for the presence of pesticide residues, while at the same time improving analytical methods and expanding the range of pesticides determined. This includes the gas chromatography method with an electronic absorption detector for the determination of eighteen organochlorine pesticide compounds in meat and meat products, fatty tissue,
milk and milk products, fish and eggs. VIRSVB analyzes between 250-300 samples annually, most of which are samples of imported fresh meat.

Heavy metals
Heavy metals are food contaminants that migrate into food and animal feed mainly from the environment. Toxic metals include metals that are not biogenic, i.e. they have exclusively toxic effects, namely: lead (Pb), mercury (Hg), cadmium (Cd), arsenic (As), thallium (Tl) and uranium (U).
Some heavy metals are essential for living organisms (biogenic), such as zinc (Zn), iron (Fe),
molybdenum (Mo), manganese (Mn), cobalt (Co) and selenium (Se). Significant amounts of metals in feed, and consequently in feed mixtures, can be the result of agricultural or industrial production methods or the result of deliberate misuse. Sources of heavy metals are in mines, metal foundries, organic and mineral fertilizers and municipal waste.
Prolonged exposure to high concentrations of these elements may increase the risk of developing
cancer and nervous system disorders, as well as impaired immune function. International
The International Agency for Research on Cancer (IARC) has classified cadmium and arsenic as Group 1 carcinogens for humans. Maximum permitted levels for certain contaminants in food are prescribed by the Regulation on Maximum Permitted Levels for Certain Contaminants in Food (Official Gazette of the Republic of Srpska 32/21 and Official Gazette of BiH 68/14, 79/16, 9/17, 84/18), and in animal feed by the Regulation on Undesirable Substances in Animal Feed (Official Gazette of BiH 72/11, 70/16).
If residue levels exceed the maximum permitted amounts, due to cumulative effects in the human body, which are a consequence of long-term food consumption, adverse health effects occur (neurotoxicity, chemotoxicity, nephrotoxicity), and therefore it is the obligation of all livestock farmers and meat processors to reduce the possibility of contamination to the lowest possible extent.
Since 2013, VIRSB has been implementing methods for the analysis of heavy metals in food and animal feed, namely: lead, cadmium, arsenic and mercury. On average, around 1,000-1,300 samples are analyzed annually for the presence of heavy metals, of which the majority are samples of imported fresh meat.

VIRSVB performs radiological tests using the HPGe Gamma Spectrometer System (P-type detector) GCDX-50190 instrument, manufactured by "Baltic Scientific Instruments" Lithuania. This instrument is used to test samples for radionuclides that are gamma emitters, of which
The most important living things are Cs-134, Cs-137, Sr-89 and I-131.
The VIRSB conducts testing for Cs-134 and Cs-137 in food, feed, water and environmental samples. In order to protect the health of the population from possible contamination, testing for the presence of radionuclides, especially Cs-137 in baby food, milk and dairy products is of great importance. Testing is carried out according to the following regulations:
– Rulebook on limits of radionuclides in food, animal feed, medicines, general purpose items, construction materials and other goods placed on the market (Official Gazette of BiH 54/14),
– Rulebook on maximum permitted quantities for certain contaminants in food (Official Gazette of BiH 68/14).

VIRSVB, as part of its regular activities, carries out sampling activities. Sampling is performed according to the request of competent authorities (inspection, ministry, etc.) and users of services, needs and requirements of projects, professional and scientific works, etc. Sampling is performed by VIRSVB staff at the Banja Luka and Bijeljina locations.
Sampling from service users, with whom there is a concluded cooperation agreement, is performed in accordance with the monthly plan, which is formed in agreement with the users.
On an annual level, VIRSVB conducts sampling of about 16,000 samples. VIRSVB samples the following types of samples: 
- samples of food, animal feed, swabs of surfaces, carcasses and water for microbiological tests,
- samples of raw milk,
- samples for testing residues and contaminants,
- samples for heavy metal testing,
– samples for pesticide testing,
-samples of meat and meat products, milk and milk products, honey and animal feed for physicochemical testing,
– samples for testing for infectious animal diseases (pathology, bacteriology, virology, serology, molecular diagnostics).

Since 2017, VIRSVB, in cooperation with the Czech Development Agency, has been participating in a project whose main purpose is to improve the food safety system in Bosnia and Herzegovina. By actively participating in the realization of one part of this project called "Training on improving implementation of national Salmonella control program - Laboratory part", VIRSVB is working on the establishment of interlaboratory
comparative study for Salmonella spp. from the stage of primary production, in order to strengthen the capacities of veterinary laboratories. This implies the organization of proficiency testing, which involves evaluating the performance of participants according to pre-established criteria using inter-laboratory comparisons. The organization of proficiency testing represents a great challenge and carries risks in terms of providing capacities and resources, both technical and personal, and ensuring the validity of the results. During seven years, VIRSVB personnel underwent several trainings, both in the Czech Republic and in the Republic of Srpska, all in accordance with the recommendations of the European Commission, for improvements in this area.
Five veterinary laboratories are participating in the proficiency test for Salmonella spp., which has been organized by VIRSVB since 2019, at the BiH level. Participation is optional, on a voluntary basis. VIRSVB organizes proficiency testing according to the BAS EN ISO/IEC 17043:2011 standard 
Conformity assessment - General requirements for proficiency testing, for the detection and serotyping of Salmonella spp. These laboratories have participated in this proficiency test for the fifth year in a row and demonstrate good laboratory practice when it comes to this type of test.

During the sale of veterinary medical products (VMP), it is very important to provide high-quality and safe VMP, as a product of special importance for the protection of the health of animals, as well as people, through the consumption of food of animal origin. VMPs can only be placed on the market if they do not endanger the health and safety of the target animals.
Based on the Law on Veterinary Medical Products (Official Gazette of Republika Srpska 71/12), VIRSVB issues an opinion on veterinary medical products imported into Republika Srpska, i.e. BiH. VIRSVB evaluates veterinary-medical
products and gives an opinion on their quality, harmlessness and effectiveness. About 40 opinions are issued annually.

VIRSVB distributes vaccination ear tags for small ruminants and vaccines against brucellosis, according to the distribution plan of the Ministry of Agriculture, Forestry and Water Management of the Republic of Srpska and the Veterinary Office of Bosnia and Herzegovina.
The Veterinary Office of Bosnia and Herzegovina prescribes the method of carrying out vaccination against brucellosis of small ruminants, based on which the vaccine and ear tags are distributed. The vaccine in question is OCUREV Rev 1, it is prepared for conjunctival application.
Marking of ear tags is carried out in accordance with the provisions of the Rulebook on marking and movement of animals in BiH (Official Gazette of BiH 3/10, 79/10, 25/11, 103/11, 41/12, 52/15, 2/2016, 66/16).
Дистрибуција ушних маркцица и вакцина ветеринарским организацијама врши се према плану који доноси Министарство пољопривреде, шумарства и водопривреде Републике Српске.
VIRSVB annually distributes about 190,000 doses of the vaccine against brucellosis and the same number of ear tags.

Hunting organizations represent an important factor in controlling the health of wild animals throughout the hunting grounds of Republika Srpska.
The significant role of hunters was also confirmed after the appearance of African swine fever in some parts of the Republic of Srpska. A quick reaction, which included the finding of dead wild boars and technical support to doctors of veterinary medicine during sampling and
rehabilitation of the terrain, was extremely important for effective disease control. By properly removing infectious material from the hunting ground, the risk of spreading the infectious disease to wider areas and even to domestic pig farms was reduced. The Hunting Association of the Republika Srpska and VIRSVB experts conducted educations throughout the Republika Srpska about this dangerous disease.No less significant role of hunters was in the monitoring of the effectiveness of vaccination of foxes against rabies in the territory of Republika Srpska. Thanks to the persistence of certain hunting organizations, a significant number of shot foxes were submitted for analysis.
Друге болести од значаја које су дијагностиковане у Републици Српској захваљујући сарадњи с ловцима јесу лажно бјеснило код ловачких паса, хемонхоза код срнеће дивљачи, синдром прољетног пролива срна, дикроцелиоза медвједа, токсоплазмоза медвједа итд.
The fruit of good cooperation between the hunting and veterinary sectors is an extensive study of parasitic diseases of wild boars and chamois, which was co-financed by the Hunting Association of the Republic of Srpska. Pioneering scientific and professional data for Republika Srpska and BiH were obtained. On the basis of coprological examinations of wild boar feces samples, the following parasitic elements were determined: eggs of the genus Metastrongylus spp., eggs of gastrointestinal strongylids, eggs of the species Ascaris suum, eggs of the species Gnathostoma hispidum, eggs of spirocercids, eggs of the genus Capillaria spp., eggs of the genus Trichuris suis and coccidia oocysts. About 75 samples of shot wild boars and four chamois were examined. Pathomorphological examination of wild boar organs revealed the following parasites:
Macracanthorhynchus hirudinaceus, Ascarops strongylina, Physocephalus sexalatus, Globocephalus spp., Metastrongylus spp., Trichuris suis and Cysticercus tenucollis. The most abundant species were Metastrongylus spp. and Trichuris suis. Other parasites were found in a smaller number of examined samples. In chamois, from the territory of the municipalities of Foča and Višegrad, the following endoparasites were detected: gut and abomasal strongylides (Haemonchus spp., Ostertagia spp., Marshallagia
spp., Trichostrongylus spp. Nematodirus spp.), Moniezia spp., Protostrongylidae and Eimeria spp. The following species and genera of ectoparasites were identified: Ixodes ricinus, Haemaphysalis spp. and Lipoptena cervi. Protostrongylids (lungworms) and the large abomasal nematode H. Contortus are an obvious health problem in chamois.
These data indicate that the Hunting Association of the Republic of Srpska is one of the key partners of VIRSVB in the fight against infectious and parasitic diseases in wild animals.
Currently, the publication "The most important infectious and parasitic diseases of game in the hunting grounds of the Republic of Srpska" is being prepared, which is intended for hunters, doctors of veterinary medicine and other users of the hunting grounds of the Republic of Srpska.

Banja Luka ćevap is an authentic specialty with a long tradition on the Balkan Peninsula, especially famous in Banja Luka. According to some records, Banja Luka ćevap has been served for more than 100 years.
Instead of the usual roll, the Banja Luka kebab is characterized by its tile shape, which consists of four joined rolls. This shape allows more fat to be retained, making it particularly juicy when consumed. In addition, Banja Luka bun, which is traditionally prepared only in this city, completes this specialty with its characteristic taste and texture.
The key elements that determine the traditional properties of the Banja Luka kebab include the composition of the kebab mixture, its shape and specific sensory properties. The Banja Luka bun is an indispensable part of this traditional specialty, without which the Banja Luka ćevap would be incomplete. It is prepared according to a traditional recipe and method and is not available on the market outside of Banja Luka. In addition to these key elements, the specifics related to the Banja Luka kebab are that the bun is baked exclusively in old ovens, the kebab is eaten hot, with the fingers, using a wooden toothpick, and the kebab is served on a plate, with a napkin and a wooden toothpick stuck through the napkin into the bun, served with red onion cut into leaves, previously salted, hot peppers, pepperoni and pepper. The interest of both domestic and foreign tourists in Banja Luka ćevap is constantly growing, which is confirmed by the festivals that have been organized in the city of Banja Luka in recent years. These festivals are dedicated to preserving the tradition of preparing and enjoying this unique traditional specialty from the coast of Vrbas. In order to popularize this product, as well as to promote the town's tourism, it was decided to start the registration procedure for Banja Luka ćevap as a guaranteed traditional product.
During 2023, at the initiative of the City of Banja Luka, VIRSVB participated in the project of registering Banja Luka ćevap as a guaranteed traditional product. The VIRSVB representative was a member of the expert team for the development of the Banja Luka ćevap specification, which included representatives of the Faculty of Technology of the University of Banja Luka, the Association for the Preservation of the Guaranteed Tradition of the Banja Luka ćevap specialty, and the City of Banja Luka. The entire procedure was carried out in accordance with the Rulebook on quality systems for food products (Official Gazette of BiH 90/18), after which on 22.03.2024 an application for registration was submitted to the Agency for Food Safety of BiH, and the registration was completed at the beginning of October 2024. VIRSVB is, for
потребе регистрације, урадио физичко-хемијска испитивања узорака Бањалучког ћевапа, те у
сарадњи с Технолошким факултетом Универзитета у Бањој Луци, урадио опис производа, опис методе производње и дефинисање кључних елемената који одређују традиционална својства производа.