Blood metabolic and hematology parameters and survivorship in mice after application of the rabies challenge virus standard in vaccine potency test

Background: Rabies virus can cause intensive and lethal infection of the central nervous system (CNS) in animals andhumans. Metabolic examinations are conducted at the cerebrospinal fluid (CSF), and it has been found that many metabolicchanges occur during RABV infection. However, although it is a neurotropic virus, it can cause damage to extraneuraltissues - lungs, heart, kidneys and liver. This study aimed to determine differences in metabolic, endocrinology and hematologic parameters in blood of mice after application of rabies challenge with virus standard 27 strain (CVS-27).Materials, Methods & Results: This study included 30 survived, and 30 dead mice that were part of the standard procedureof NIH (National Institute of Health) test in Pasteur Institute in Novi Sad. Tests were performed in the following order: twogroups of mice were vaccinated in a 7 day period with different dilutions of standard vaccine and the examined vaccine.Seven days after the last vaccination, immunized animals and animals in the control group received test virus CVS-27.Blood samples were collected from a heart puncture. Differences in hematologic and biochemical parameters were determined by t-test. Due to a high number of blood parameters, we performed a joint analysis of multiple dependent variables.Higher pH value and higher concentrations of glucose, cholesterol, lactate dehydrogenase (LDH), creatine kinase (CK),albumin, urea, creatinine, α-amylase, magnesium (Mg), nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHB)and lactate were noted in dead mice. Higher granulocytes and mean platelet volume (MPV) were noted in mice whichdied, but also reduced lymphocytes, erythrocytes, haemoglobin, hematocrit and platelets count. Higher values of insulin,cortisol and HOMA-IR (homeostatic model assessment insulin resistance) were noted in the group of dead mice comparedto the surviving one. Reduced QUICKI...

Blood metabolic and hematology parameters and survivorship in mice after application of the rabies challenge virus standard in vaccine potency test

Background: Rabies virus can cause intensive and lethal infection of the central nervous system (CNS) in animals andhumans. Metabolic examinations are conducted at the cerebrospinal fluid (CSF), and it has been found that many metabolicchanges occur during RABV infection. However, although it is a neurotropic virus, it can cause damage to extraneuraltissues - lungs, heart, kidneys and liver. This study aimed to determine differences in metabolic, endocrinology and hematologic parameters in blood of mice after application of rabies challenge with virus standard 27 strain (CVS-27).Materials, Methods & Results: This study included 30 survived, and 30 dead mice that were part of the standard procedureof NIH (National Institute of Health) test in Pasteur Institute in Novi Sad. Tests were performed in the following order: twogroups of mice were vaccinated in a 7 day period with different dilutions of standard vaccine and the examined vaccine.Seven days after the last vaccination, immunized animals and animals in the control group received test virus CVS-27.Blood samples were collected from a heart puncture. Differences in hematologic and biochemical parameters were determined by t-test. Due to a high number of blood parameters, we performed a joint analysis of multiple dependent variables.Higher pH value and higher concentrations of glucose, cholesterol, lactate dehydrogenase (LDH), creatine kinase (CK),albumin, urea, creatinine, α-amylase, magnesium (Mg), nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHB)and lactate were noted in dead mice. Higher granulocytes and mean platelet volume (MPV) were noted in mice whichdied, but also reduced lymphocytes, erythrocytes, haemoglobin, hematocrit and platelets count. Higher values of insulin,cortisol and HOMA-IR (homeostatic model assessment insulin resistance) were noted in the group of dead mice comparedto the surviving one. Reduced QUICKI...(AU)

Relationship of circulating tumor necrosis factor alpha (TNF-alfa) and insulin secretion and resistance in euglycaemic dogs

Background: Insulin resistance is a state that is characterized with reduced sensitivity of peripheral tissues to insulin. Itcan be related with increased level of tumor necrosis factor alpha (TNF-α) in dogs. Insulin resistance can be evaluated byhomeostasis model assessment (HOMA-IR, HOMA-β). The aim of this study was to determine correlation of circulatingTNF-α level with insulin production and insulin resistance indexes in euglycaemic dogs.Materials, Methods & Results: Seventy dogs of normal body score were included in this study. After blood sampling levelsof glucose, insulin and TNF-α were determined and indexes HOMA-IR and HOMA-β were calculated. Three groups inaccordance to TNF-α levels were formed: the first-TNF-α 0-2.0 pg/mL, the second-TNF-α below median (2.1-17.0 pg/mL)and the third-TNF-α above median (17.1-51.8 pg/mL). Differences in insulin and glucose levels, HOMA-IR and HOMA-βwere determined in all three groups. ANOVA and posthock LSD analyses were used. Correlation between HOMA-IR andHOMA-β was determined. Linear regression between HOMA-β/HOMA-IR ratio and glucose concentration was calculated. SPSS statistical program was used (IBM). Highest insulin level was detected in the second group and the lowest wasdetected in the third group. The lowest glucose level was detected in the first group. The highest value of HOMA-β indexwas noted in the first group and it decreases with TNF-α increase. The highest HOMA-IR value was detected in the secondgroup and the lowest was in the third group. Positive correlation was noted between HOMA-IR and HOMA-β. Significantlinear correlation was noted between glucose levels in function of HOMA-β/HOMA-IR...

Prevalence of Dirofilaria immitis in dogs from shelters in Vojvodina, Serbia

Background: Dirofilaria immitis is vector borne parasite of carnivores, with zoonotic potential, endemic in many parts of the world, including Europe. The aim of this study was to determine the prevalence of Dirofilaria immitis infection in dogs from shelters, especially compared to their lifestyle. Dogs living in shelters in Serbia may be at high risk of acquiring vector borne pathogens, mainly because most of them live outside in pens and backyards, in contact with vectors. Also, dogs in shelters are not always regularly treated against ectoparasites, thus, representing an easy feeding source for the vectors. The objective of this study was to determine the prevalence of Dirofilaria immitis infection in dogs from 5 shelters in South Bačka and Central Banat districts, in Autonomous Province of Vojvodina, Northern part of Serbia. Also, the objective was to compare the relation of infection with Dirofiaria immitis with age, sex, type of keeping the animals and preventive treatment in dogs. Materials, Methods & Results: Between May 2017 and October 2019, blood samples were collected from 336 randomly selected dogs from 5 shelters in 2 districts, South Bačka and Central Banat districts, in Autonomous Province of Vojvodina, Northern part of Serbia. The epidemiological survey has been conducted with all of the dogs involved in this research. The survey was designed to collect data about sex, age, lifestyles, food type, treatment against mosquitoes with insecticides and filarioid worms with macrocyclic lactones, regular testing for Dirofilaria infections. The presence of circulating microfilariae was examined using a modified Knott’s test. For the presence of circulating adult female Dirofilaria immitis antigen, serum samples were tested by commercially available enzyme-linked immunosorbent assay, which reacts to antigen of female Dirofilaria. In total, 336 dogs were examined for the presence of Dirofilaria...

Relationship of circulating tumor necrosis factor alpha (TNF-alfa) and insulin secretion and resistance in euglycaemic dogs

Background: Insulin resistance is a state that is characterized with reduced sensitivity of peripheral tissues to insulin. Itcan be related with increased level of tumor necrosis factor alpha (TNF-α) in dogs. Insulin resistance can be evaluated byhomeostasis model assessment (HOMA-IR, HOMA-β). The aim of this study was to determine correlation of circulatingTNF-α level with insulin production and insulin resistance indexes in euglycaemic dogs.Materials, Methods & Results: Seventy dogs of normal body score were included in this study. After blood sampling levelsof glucose, insulin and TNF-α were determined and indexes HOMA-IR and HOMA-β were calculated. Three groups inaccordance to TNF-α levels were formed: the first-TNF-α 0-2.0 pg/mL, the second-TNF-α below median (2.1-17.0 pg/mL)and the third-TNF-α above median (17.1-51.8 pg/mL). Differences in insulin and glucose levels, HOMA-IR and HOMA-βwere determined in all three groups. ANOVA and posthock LSD analyses were used. Correlation between HOMA-IR andHOMA-β was determined. Linear regression between HOMA-β/HOMA-IR ratio and glucose concentration was calculated. SPSS statistical program was used (IBM). Highest insulin level was detected in the second group and the lowest wasdetected in the third group. The lowest glucose level was detected in the first group. The highest value of HOMA-β indexwas noted in the first group and it decreases with TNF-α increase. The highest HOMA-IR value was detected in the secondgroup and the lowest was in the third group. Positive correlation was noted between HOMA-IR and HOMA-β. Significantlinear correlation was noted between glucose levels in function of HOMA-β/HOMA-IR...(AU)

Correlation between different Helicobacter morphotypes and histological changes in pig gastric mucosa

Background: Two distinct morphologic types of bacteria which belong to the Helicobacter species, have been described in pigs: once or twice curved Helicobacter pylori-like bacteria (HLO) and the multicoiled, Gastrospirillum-like bacteria (GLO). The objective of this study was to determine the incidence of Helicobacter spp. using modified Giemsa stain and to define the relationship between presence of Helicobacter spp. and histopathological changes of gastric mucosa in pigs.Materials, Methods & Results: A total of 120 pig stomachs (60 from intensive and 60 from extensive breeding) were enrolled in this study and 240 fragments of fundic and pyloric mucosa were taken for histopathological examination. By modified Giemsa staining of gastric mucosa, Helicobacter-like organisms were confirmed in 4/60 (6.67%) of pigs in intensive and 5/60 (8.33%) of pigs in extensive breeding. The incidence of tightly spiral shaped Gastrospirillum-like organisms in pigs of intensive and extensive breeding were 5/60 (8.33%) and 9/60 (15%), respectively. The severity of gastritis was scored to the Sydney System with some modifications. There was no significant difference between HLO-positive and HLO-negative fundic mucosa in pigs of both breeding systems. In contrary, there was significant difference between HLO-positive and HLO-negative pyloric mucosa of pigs in intensive (P < 0.001) and in extensive breeding (P < 0.05). In intensive breeding, there was significant difference between GLO-positive and GLO-negative fundic mucosa (P < 0.05), while there was no significant difference between GLO-positive and GLO-negative pyloric mucosa.Discussion: The prevalence of Helicobacter spp. bacteria were in coherence with literature data. The higher prevalence of both morfological type of bacteria, were considered in pigs in extensive breeding. the hygienic conditions and managment factors in pigs farm are the possible impact for higher bacterial transmission.[...]

Correlation between different Helicobacter morphotypes and histological changes in pig gastric mucosa

Background: Two distinct morphologic types of bacteria which belong to the Helicobacter species, have been described in pigs: once or twice curved Helicobacter pylori-like bacteria (HLO) and the multicoiled, Gastrospirillum-like bacteria (GLO). The objective of this study was to determine the incidence of Helicobacter spp. using modified Giemsa stain and to define the relationship between presence of Helicobacter spp. and histopathological changes of gastric mucosa in pigs.Materials, Methods & Results: A total of 120 pig stomachs (60 from intensive and 60 from extensive breeding) were enrolled in this study and 240 fragments of fundic and pyloric mucosa were taken for histopathological examination. By modified Giemsa staining of gastric mucosa, Helicobacter-like organisms were confirmed in 4/60 (6.67%) of pigs in intensive and 5/60 (8.33%) of pigs in extensive breeding. The incidence of tightly spiral shaped Gastrospirillum-like organisms in pigs of intensive and extensive breeding were 5/60 (8.33%) and 9/60 (15%), respectively. The severity of gastritis was scored to the Sydney System with some modifications. There was no significant difference between HLO-positive and HLO-negative fundic mucosa in pigs of both breeding systems. In contrary, there was significant difference between HLO-positive and HLO-negative pyloric mucosa of pigs in intensive (P < 0.001) and in extensive breeding (P < 0.05). In intensive breeding, there was significant difference between GLO-positive and GLO-negative fundic mucosa (P < 0.05), while there was no significant difference between GLO-positive and GLO-negative pyloric mucosa.Discussion: The prevalence of Helicobacter spp. bacteria were in coherence with literature data. The higher prevalence of both morfological type of bacteria, were considered in pigs in extensive breeding. the hygienic conditions and managment factors in pigs farm are the possible impact for higher bacterial transmission.[...](AU)

Analysis of different diagnostis methods of influenza in horses

Background: Equine Influenza is a serious, acute respiratory illness with characteristical clinical signs. The disease is caused by family of Orthomyxoviridae, genera Influenza virus A by two subtypes H7N7 and H3N8. Currently, there is believe that H7N7 has been replaced as a predominant subtype with the H3N8. Horse infection with influenza virus can be detected by serological tests on paired sera using HI test. Commercial rapid tests could be used for the detection of influenza virus. Recently it is widely use a PCR method as fast and more specific methods. Materials, Methods & Results: Fifty horses and one pony, age between one and 22 years have been included in experiment. Horses were of different race, sex, and age and vaccination status. Ten out of total 51 (10/51) have been regularly vaccinated against EI. Prior to initiation of these study epidemiological survey has been performed. The clinical examination has been followed by blood sampling for blood cell and serum extraction. The serums were evaluated by HI method. Nasal swabs are taken from both nostrils twice, one was frozen for virus detection by RT-qPCR while another was used for detection of EI virus by Directi-gen FLU A rapid test. Analysis of titers of antibody reveled that 7 horses (14%) had specific antibodies (IgG) against subtype H7N7, while 9 horses (18%) had specific antibodies against H3N8 [...](AU)

Analysis of different diagnostis methods of influenza in horses

Background: Equine Influenza is a serious, acute respiratory illness with characteristical clinical signs. The disease is caused by family of Orthomyxoviridae, genera Influenza virus A by two subtypes H7N7 and H3N8. Currently, there is believe that H7N7 has been replaced as a predominant subtype with the H3N8. Horse infection with influenza virus can be detected by serological tests on paired sera using HI test. Commercial rapid tests could be used for the detection of influenza virus. Recently it is widely use a PCR method as fast and more specific methods. Materials, Methods & Results: Fifty horses and one pony, age between one and 22 years have been included in experiment. Horses were of different race, sex, and age and vaccination status. Ten out of total 51 (10/51) have been regularly vaccinated against EI. Prior to initiation of these study epidemiological survey has been performed. The clinical examination has been followed by blood sampling for blood cell and serum extraction. The serums were evaluated by HI method. Nasal swabs are taken from both nostrils twice, one was frozen for virus detection by RT-qPCR while another was used for detection of EI virus by Directi-gen FLU A rapid test. Analysis of titers of antibody reveled that 7 horses (14%) had specific antibodies (IgG) against subtype H7N7, while 9 horses (18%) had specific antibodies against H3N8 [...]

Importance of breeding pigs in the spread of Salmonella on farms

Background: Pigs infected with Salmonella represent a high risk for contamination and cross-contamination of carcasses with Salmonella from feces, tonsils and mesenteric lymph nodes, which occurs on the slaughter line. The environment itself represents a potential source of infection, as it is Salmonella infection of other individuals in the herd (sows, boars, piglets, fattener pigs). One of the leading sources of infection is contaminated feed for pigs, or its ingredients. If an infection occurs at any stage and category of production, those individual animals can become a potential source of infection for all others on farm. Materials, Methods & Results: This study examines the importance of breeding categories of pigs in the spread of salmonellosis in piglets and fattener pigs on seven farrow-to-finish farms in Serbia. From each farm, 30 sows, 121 boars and 90 piglets were tested from the weaning to the time of dispatch to the slaughterhouse. Pigs were examined for the presence of Salmonella-specific antibodies in blood sera and the presence of Salmonella in feces. Pigs feed on the farms were also examined for the presence of Salmonella. The identification of serotypes was conducted according to the Kauffmann-White scheme. For antimicrobial sensitivity and resistance testing, isolates were tested by disc diffusion against a panel of 11 antimicrobials. The numbers...(AU)

Importance of breeding pigs in the spread of Salmonella on farms

Background: Pigs infected with Salmonella represent a high risk for contamination and cross-contamination of carcasses with Salmonella from feces, tonsils and mesenteric lymph nodes, which occurs on the slaughter line. The environment itself represents a potential source of infection, as it is Salmonella infection of other individuals in the herd (sows, boars, piglets, fattener pigs). One of the leading sources of infection is contaminated feed for pigs, or its ingredients. If an infection occurs at any stage and category of production, those individual animals can become a potential source of infection for all others on farm. Materials, Methods & Results: This study examines the importance of breeding categories of pigs in the spread of salmonellosis in piglets and fattener pigs on seven farrow-to-finish farms in Serbia. From each farm, 30 sows, 121 boars and 90 piglets were tested from the weaning to the time of dispatch to the slaughterhouse. Pigs were examined for the presence of Salmonella-specific antibodies in blood sera and the presence of Salmonella in feces. Pigs feed on the farms were also examined for the presence of Salmonella. The identification of serotypes was conducted according to the Kauffmann-White scheme. For antimicrobial sensitivity and resistance testing, isolates were tested by disc diffusion against a panel of 11 antimicrobials. The numbers...

Seroepidemiological research of canine monocytic ehrlichiosis in the autonomous province of Vojvodina, Serbia

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a 'new' infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs' lifestyles and information on whether the dogs had been abroad. An overall of 62 ticks were collected from the bodies of these dogs. By applying the screening iELISA test, the seroprevalence of class G specific antibodies was determined with dogs to E. canis antigens in 25%, while 75% of dogs were negative to the presence of specific antibodies of IgG class to E. canis. Out of 25 seropositive dogs, with antibodies to E. canis discovered through iELISA test, 12 (48%) dogs were in contact with ticks for the first time, while the repeated contact with ticks was experienced by 9 (36%) dogs. For 4 (16%) dogs, the owners negated that their dog had been bitten by a tick. Upon the application of a confirming IFAT, a seroprevalence was determined of specific class G antibodies with dogs to E. canis antigens in 16%, while 84% were negative to the presence of specific antibodies of IgG class antibodies to E. canis. Out of 16 seropositive dogs, in which antibodies were discovered with IFAT to E. canis, 9 (56%) dogs had been in contact with ticks for the fi rst time, while 4 dogs (25%) had repeated contacts with ticks. For 3 dogs (19%), the owners negated that ticks had bitten their dogs. Through a statistical processing of the results gained from the applied immunodiagnostic tests, a moderate agreement (Kappa value of 0.48) was determined. In the group of dogs that had been bitten by a tick, three types were determined: Dermacentor marginatus, Rhipicephalus sanguineus and Ixodes ricinus. Overall 35 ticks of Dermacentor marginatus ticks were replaced from the dogs' bodies just as were 15 Rhipicephalus sanguineus ticks and 12 Ixodes ricinus ticks. Discussion: The determined values of the seroprevalence in our country (iELISA - 25% and IFAT - 16%) are in harmony with the acquired values in other countries with warm climate. The values of seroprevalence determined in this research point to the fact that this disease must be taken into consideration in everyday health protection of dogs, especially after a registered tick bite within a differential diagnostics procedure.

Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia / Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o

Findings of antibodies to Mycoplasma suis on swine farms in Serbia / Findings of antibodies to Mycoplasma suis on swine farms in Serbia

Background: Hemoplasmas are eperythrocytic procaryotes, including Mycoplasma species which were recently transferred from the genera Eperythrozoon and Haemobartonella, based on their similarity of the 16S rRNA sequences, and newly identified hemotropic mycoplasmas. Eperythrozoonosis of pigs is caused by the haemotrophic bacterium M. suis, and the disease has a worldwide distribution. The disease manifests as a severe and often fatal acute febrile icteroanemia, mainly in piglets, pregnant sows before parturition and fattening pigs exposed to stress. The aim of this study was to determine the seroprevalence of IgG to M. suis in swine populations in Serbia, using a Western Blot test with the recombinant protein MSG1. Materials, Methods & Results: Four farms were chosen to represent the main swine-producing geographic regions of Serbia, including South Baka District (farms 1 and 2), North Baka District (farm 3) and Branievo District (farm 4). A total of forty-six clinically healthy pigs, age 8-20 weeks, were included in the study. Blood samples from pigs randomly selected from the four farms were collected by jugular vein puncture into serum vacutainer tubes with clot activator. After clotting at room temperature for 1 h, blood samples were centrifuged at 1500 g, for 15 min at room temperature. Sera were carefully harvested and stored at -20C until assayed. The sera samples

Background: Hemoplasmas are eperythrocytic procaryotes, including Mycoplasma species which were recently transferred from the genera Eperythrozoon and Haemobartonella, based on their similarity of the 16S rRNA sequences, and newly identified hemotropic mycoplasmas. Eperythrozoonosis of pigs is caused by the haemotrophic bacterium M. suis, and the disease has a worldwide distribution. The disease manifests as a severe and often fatal acute febrile icteroanemia, mainly in piglets, pregnant sows before parturition and fattening pigs exposed to stress. The aim of this study was to determine the seroprevalence of IgG to M. suis in swine populations in Serbia, using a Western Blot test with the recombinant protein MSG1. Materials, Methods & Results: Four farms were chosen to represent the main swine-producing geographic regions of Serbia, including South Baka District (farms 1 and 2), North Baka District (farm 3) and Branievo District (farm 4). A total of forty-six clinically healthy pigs, age 8-20 weeks, were included in the study. Blood samples from pigs randomly selected from the four farms were collected by jugular vein puncture into serum vacutainer tubes with clot activator. After clotting at room temperature for 1 h, blood samples were centrifuged at 1500 g, for 15 min at room temperature. Sera were carefully harvested and stored at -20C until assayed. The sera samples

Findings of antibodies to Mycoplasma suis on swine farms in Serbia

Background: Hemoplasmas are eperythrocytic procaryotes, including Mycoplasma species which were recently transferred from the genera Eperythrozoon and Haemobartonella, based on their similarity of the 16S rRNA sequences, and newly identified hemotropic mycoplasmas. Eperythrozoonosis of pigs is caused by the haemotrophic bacterium M. suis, and the disease has a worldwide distribution. The disease manifests as a severe and often fatal acute febrile icteroanemia, mainly in piglets, pregnant sows before parturition and fattening pigs exposed to stress. The aim of this study was to determine the seroprevalence of IgG to M. suis in swine populations in Serbia, using a Western Blot test with the recombinant protein MSG1. Materials, Methods & Results: Four farms were chosen to represent the main swine-producing geographic regions of Serbia, including South Backa District (farms 1 and 2), North Backa District (farm 3) and Branicevo District (farm 4). A total of forty-six clinically healthy pigs, age 8-20 weeks, were included in the study. Blood samples from pigs randomly selected from the four farms were collected by jugular vein puncture into serum vacutainer tubes with clot activator. After clotting at room temperature for 1 h, blood samples were centrifuged at 1500 g, for 15 min at room temperature. Sera were carefully harvested and stored at -20°C until assayed. The sera samples were tested by the Western Blot test with recombinant protein MSG1 (p40). Production of recombinant protein MSG1 (p40), dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting were performed as previously described by Hoelzle. Specific IgG antibodies to MSG1 of M. suis were identified in 20 of the 46 samples tested, giving a total seroprevalence of 43.47%. The M. suis-specific antibody response was detected in pigs from all tested farms, within farm seroprevalences of 54.54% on farm 1; 27.27% on farm 2; 36.36% on farm 3 and 53.84% on farm 4. Discussion: Reports on the prevalence of swine infected with Mycoplasma suis in other countries are rare, but have been communicated for USA, Brazil, Japan, Portugal, China and Germany. Previously published values of the prevalence of the pigs infected with haemoplasmas in Serbia, determined through a microscopic examination of the peripheral blood smear according to Giemsa was 39% and with Acridine orange was 47%. These prevalences are similar to the overall prevalence found in this study (43.47%). In this present study, the prevalence of M. suis in the farm pigs studied in Serbia (43.47%) was higher than the serological evidence of the infection reported in pigs in the USA, Brazil and Japan, but lower than the prevalence in sows in Portugal and China. In this present study, the prevalence of M. suis on the studied pig farms in Serbia (43.47%) was more similar to results obtained in Germany, where M. suis infections were detected in 79 out of 196 pig farms (40.3%) by employing a quantitative real-time LightCycler PCR. Also, M. suis was detected in 36 out of 359 wild German boars (10.03%) with similar methodology. Our seroprevalence of anti-M. suis IgG in farmed pigs is probably higher than that in wild boar because of the conditions of intensive breeding on pig farms, but perhaps this may also be partly due to the different geographic locality, and/or to different analytical methods. This overall determined seroprevalence of 43.47% from a small sample within a relatively small area suggests a signifi cantly higher presence of infection on pig farms in Serbia, and hence, significantly more economic losses in pig production than could be expected based on the worldwide reports of M. suis prevalence.

A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cow's milk

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed. Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37ºC in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by double CAMP and plasma coagulation tube test. All 14 isolates developed a synergistic haemolysis with Rhodococcus equi (ATCC 6939) and inverse CAMP phenomenon with Staphylococcus aureus and coagulated rabbit plasma. Final diagnosis was confirmed using API Coryne V 2.0 and software program by BioMerieux1, revealing an identity rate of 99.9%, accuracy rate T = 1, test count = 0. Discussion: The first fourteen isolates of Corynebacterium ulcerans have been identified in our country, on the basis of a diagnostic protocol that is proposed in this paper. In our experience double CAMP test, rabbit plasma coagulation, catalase, oxidase tests and selected biochemical parameters, are sufficient as a diagnostic minimum. In the diagnostics of bacterial agents in cow mastitis, the attention of a bacteriologist is mostly limited to most widespread agents of mastitis, the isolation of which is mandatory pursuant to national legislation (Staphylococcus aureus and Streptococcus agalactiae). A more important reason for "missing" Corynebacterium ulcerans in the diagnosis is its colonial morphology that could resemble organisms of the genus Staphylococcus. Complex and expensive diagnostic procedure that is not available to most laboratories is also responsible for the small number of reports of isolation C. ulcerans. Furthermore, in routine work C. ulcerans could be misidentified with Staphylococcus intermedius, because of cultural similarity, positive plasma coagulation tube test and absence of manitol fermentation of both species. This paper is a report on isolation and identification of Corynebacterium ulcerans from milk of cows with mastitis, as well as a suggestion of a diagnostic protocol available for routine work in most veterinary microbiology laboratory. Therefore we suggest as the diagnostic protocol double CAMP test to be used as a complementary method to rabbit plasma coagulation tube test.

A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk / A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Findings of antibodies to Mycoplasma suis on swine farms in Serbia / Findings of antibodies to Mycoplasma suis on swine farms in Serbia

Background: Hemoplasmas are eperythrocytic procaryotes, including Mycoplasma species which were recently transferred from the genera Eperythrozoon and Haemobartonella, based on their similarity of the 16S rRNA sequences, and newly identified hemotropic mycoplasmas. Eperythrozoonosis of pigs is caused by the haemotrophic bacterium M. suis, and the disease has a worldwide distribution. The disease manifests as a severe and often fatal acute febrile icteroanemia, mainly in piglets, pregnant sows before parturition and fattening pigs exposed to stress. The aim of this study was to determine the seroprevalence of IgG to M. suis in swine populations in Serbia, using a Western Blot test with the recombinant protein MSG1. Materials, Methods & Results: Four farms were chosen to represent the main swine-producing geographic regions of Serbia, including South Baka District (farms 1 and 2), North Baka District (farm 3) and Branievo District (farm 4). A total of forty-six clinically healthy pigs, age 8-20 weeks, were included in the study. Blood samples from pigs randomly selected from the four farms were collected by jugular vein puncture into serum vacutainer tubes with clot activator. After clotting at room temperature for 1 h, blood samples were centrifuged at 1500 g, for 15 min at room temperature. Sera were carefully harvested and stored at -20C until assayed. The sera samples

Background: Hemoplasmas are eperythrocytic procaryotes, including Mycoplasma species which were recently transferred from the genera Eperythrozoon and Haemobartonella, based on their similarity of the 16S rRNA sequences, and newly identified hemotropic mycoplasmas. Eperythrozoonosis of pigs is caused by the haemotrophic bacterium M. suis, and the disease has a worldwide distribution. The disease manifests as a severe and often fatal acute febrile icteroanemia, mainly in piglets, pregnant sows before parturition and fattening pigs exposed to stress. The aim of this study was to determine the seroprevalence of IgG to M. suis in swine populations in Serbia, using a Western Blot test with the recombinant protein MSG1. Materials, Methods & Results: Four farms were chosen to represent the main swine-producing geographic regions of Serbia, including South Baka District (farms 1 and 2), North Baka District (farm 3) and Branievo District (farm 4). A total of forty-six clinically healthy pigs, age 8-20 weeks, were included in the study. Blood samples from pigs randomly selected from the four farms were collected by jugular vein puncture into serum vacutainer tubes with clot activator. After clotting at room temperature for 1 h, blood samples were centrifuged at 1500 g, for 15 min at room temperature. Sera were carefully harvested and stored at -20C until assayed. The sera samples

A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk / A proposal of a diagnostic protocol for isolation of Corynebacterium ulcerans from cows milk

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Background: Literature about presence Corynebacterium ulcerans in milk samples from cows with mastitis is rare and in the literature there are only a few reports. In this study the isolation and identification of Corynebacterium ulcerans from mastitis in dairy cows were done. Also, optimization of diagnostic protocols to identify Corynebacterium ulcerans was performed.Materials, Methods & Results: The investigation was performed at the cattle farm that is characterized by closed housing system diary Holstein-Friesian cows during an outbreak of acute mastitis. Milk samples from 298 lactating cows were collected in sterile sampling tubes. Before the collection of quarter milk samples, the udder was thoroughly cleaned with soap and water and rubbed to dry. All collected milk samples were examined for mastitis using California mastitis test, which was carried out by the method first described by Schalm and Noorlander. Equal volumes (5 mL) of commercial CMT reagent and quarter milk were mixed and the changes in milk fluidity and viscosity were observed. Sample portions (0.1 mL each) were inoculated on 10% sheep blood agar, Endo agar and Sabouraud agar as well as on thioglycolate medium and nutrient broth. Primary plates were incubated for 3 days at 37o C in aerobic conditions. Cultural, morphological and conventional biochemical testing was done. The survey was complemented by d

Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia / Seroepidemiological research of canine monocytic ehrlichiosis in the Autonomous Province of Vojvodina, Serbia

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o

Background: Monocytic ehrlichiosis is an important disease in canine pathology and is present worldwide and has a potentially lethal outcome. This infection as a new infective disease of dogs and people and it has not been sufficiently researched, especially from the aspect of clinical picture, pathogenesis, diagnostics and differential diagnostics. The aims of this seroepidemiological research of the E. canis infection with dogs are to determine the values of G class antibodies prevalence to E. canis antigens, also to determine a agreement of positive serological findings between two immunodiagnostic tests (iELISA and IFAT), and to make an overview of the epidemiological characteristics of this canine infection. Materials, Methods & Results: One hundred dogs were involved in the seroepidemiological research of the E. canis infection. These dogs were from the City of Novi Sad area and its outskirts, Autonomous Province of Vojvodina, Serbia. The epidemiological survey of the closed type has been used in the research. The survey has been conducted with all of the dogs involved in this research. Apart from identifying veterinary practices and the patients (breed, sex, age), the survey was designed to collect data about the tick bites (first bite or a repeated one); the number of the removed ticks; the dogs lifestyles and information on whether the dogs had been abroad. An o