Trypanosomosis is an important disease of dromedary camels caused by the pathogenic protozoan Trypanosoma evansi. This study aimed to compare three different tests for its diagnosis in this species: conventional microscopy, the card agglutination test for trypanosomosis/T. evansi (CATT/T. evansi) and real-time PCR. Whole blood and serum samples collected from 77 dromedary camels of Abu Dhabi, United Arab Emirates, were analysed with the test methods stated. Statistical analysis was done using McNemar’s chi-squared test, and Cohen’s kappa index (κ) was calculated. We obtained results with positivity of 18% (14/77) by microscopy, 22% by CATT (17/77) and 60% (46/77) by real-time PCR, with the chain reaction detecting at a respectively three- and two-fold greater rate than the other techniques. Analysis of the data revealed a relative sensitivity of 30.4% and 37.0% for microscopy and CATT, respectively, compared to real-time PCR. The difference between the real-time PCR’s sensitivity and those of the other methods was statistically significant, with X² values of 30.03 and 20.1, respectively (df = 1 and P = 0.05 in both cases). Agreement of microscopy results with those of with CATT was good (κ = 0.72; 95% CI = 0.62–0.82). Cohen’s kappa index showed fair agreement of real-time PCR with microscopy (κ = 0.26; 95% CI = 0.16–0.36) whereas it was in poor agreement with CATT (κ = 0.09; 95% CI = 0.02–0.15). Real-time PCR was found to be more sensitive than microscopy and CATT.

Trypanosomosis is an important disease of dromedary camels caused by the pathogenic protozoan Trypanosoma evansi. This study aimed to compare three different tests for its diagnosis in this species: conventional microscopy, the card agglutination test for trypanosomosis/T. evansi (CATT/T. evansi) and real-time PCR.

The prevalence of Dirofilaria repens in dogs in countries bordering Slovenia ranges from 1.5% to 47.3%. The aim of this study was to estimate its prevalence in Slovenian dogs and to present the cases of dirofilariasis diagnosed in humans from 2010 to 2020. Epidemiological data were collected and blood samples were taken from 465 dogs older than one year and born in Slovenia. A real-time PCR was performed on all samples to detect filarioid DNA, and a D. repens-and D. immitis-specific real-time PCR was performed on positive samples. Blood samples from 446 dogs were tested for Dirofilaria spp. using a modified Knott’s test. Human cases were diagnosed from histological sections of excised subcutaneous nodules. Descriptive statistics were used to characterise the samples. The one-sample nonparametric chi-squared test was used to assess whether categories of a variable were equally distributed. Three dogs’ samples tested positive for D. repens using the species-specific real-time PCR, while D. immitis DNA was not detected. The modified Knott’s test was positive in two of the three PCR-positive dogs, two of which had never travelled outside Slovenia’s borders. Four human patients with D. repens dirofilariasis were diagnosed. Since their travel history was unknown, autochthonous transmission could not be confirmed. Our study demonstrated a 0.64% prevalence of D. repens infection in dogs in Slovenia. Two cases could be autochthonous.

The prevalence of Dirofilaria repens in dogs in countries bordering Slovenia ranges from 1.5% to 47.3%. The aim of this study was to estimate its prevalence in Slovenian dogs and to present the cases of dirofilariasis diagnosed in humans from 2010 to 2020.

The pseudorabies virus (PRV) gene encoding thymidine kinase (tk) is an important virulence-associated factor. Attenuation of PRV in susceptible animals is a frequent result of tk deletion. The aim of the study was to assess the pathogenicity of tk-deleted PRV in rats. Sprague Dawley rats were infected with the tk-deleted PRV strain SuHV-1 ΔTK:247via intranasal or intramuscular inoculation. PRV loads in ten tissues from dead and euthanised rats were determined using real-time PCR. Infection with SuHV-1 ΔTK:247 could cause death in rats. The 50% lethal dose (LD₅₀) of SuHV-1 ΔTK:247 via intranasal inoculation was 10³.¹⁶ TCID₅₀ in rats. Intramuscular inoculation required a higher dose of SuHV-1 ΔTK:247 (10⁵.⁰ TCID₅₀). A high SuHV-1 ΔTK:247 titre was observed in the trigeminal ganglia or spinal cord of dead rats. The results of this study show that rats are highly susceptible to PRV infection, and tk deletion did not completely diminish the pathogenicity of PRV in rats.

The pseudorabies virus (PRV) gene encoding thymidine kinase (tk) is an important virulence-associated factor. Attenuation of PRV in susceptible animals is a frequent result of tk deletion. The aim of the study was to assess the pathogenicity of tk-deleted PRV in rats.

Leishmaniasis is a life-threatening zoonosis of which dogs are the major reservoir and sandflies are the vectors. Until now, the prevalence of canine leishmaniasis (CanL) in the Slovenian dog population was unknown. Epidemiological data, eye swabs and blood samples were taken from 465 dogs born in Slovenia and older than one year. Commercial ELISA kits and real-time PCR were used. For ELISA-positive samples, an immunofluorescence antibody test (IFAT) was performed. Descriptive statistics were used to characterise the samples. The one-sample nonparametric chi-square test was used to test whether the categories of a variable were equally distributed. A 59.9% proportion of the recruited dogs had travelled to endemic regions and 62.1% of them had not been protected by insect repellents. Skin symptoms that might be CanL-related were described in 109 of the dogs’ histories (23.4%), inappetence and/or weight loss in 25 (5.4%), and anaemia, intermittent fever, and/or lymphadenopathy in 19 (4.1%). At the time of recruitment, all dogs were asymptomatic. All samples were PCR negative, nine (1.9%) were ELISA positive, but none were IFAT positive. Five of the nine ELISA-positive dogs were non-travellers. We conclude that the seroprevalence of canine leishmaniasis of 1.9 % in the autochthonous Slovenian dog population may pose a risk of endemic spread of the disease.

Leishmaniasis is a life-threatening zoonosis of which dogs are the major reservoir and sandflies are the vectors. Until now, the prevalence of canine leishmaniasis (CanL) in the Slovenian dog population was unknown.

Diarrhoea in growing-finishing pigs is a common problem of commercial pig farms. Among many causative factors, porcine circovirus type 2 (PCV2) is one considered an important pathogen in modern pig production. The aim of the study was to verify if PCV2 was responsible for antibiotic non-responsive diarrhoea and wasting in pigs. A total of 13 dead pigs aged between 12 and 15 weeks from three Polish farms with persistent herd symptoms suggestive of PCV2 infection were provided for evaluation. Sections of lymph nodes and intestines were analysed by in situ hybridization (ISH) for PCV2 and histopathological examination. Faeces and intestinal scrapings were tested for Lawsonia intracellularis and Brachyspira hyodysenteriae by real-time PCR and for parasitic infection by flotation and decantation. ISH and histopathological examination showed that all pigs were PCV2 systemic disease negative. Swine dysentery was confirmed by real-time PCR on two farms, and proliferative enteropathy on one farm. In histological examinations, erosions of the caecal and colonic mucosa were found, together with cysts and trophozoites of Balantidium coli. The protozoa were present in the intestinal lumen and mucosa. B. coli cysts were identified in faeces from all examined pigs. These results suggest that monitoring of B. coli infections should be an additional measure of control and prevention of gastrointestinal tract disorders in modern swine husbandry.

Diarrhoea in growing-finishing pigs is a common problem of commercial pig farms. Among many causative factors, porcine circovirus type 2 (PCV2) is one considered an important pathogen in modern pig production. The aim of the study was to verify if PCV2 was responsible for antibiotic non-responsive diarrhoea and wasting in pigs.

Tularaemia is a zoonotic disease caused by the gram-negative bacterium Francisella tularensis, which is endemic to Ukraine. The aim of this work was to provide screening of different field samples (rodent tails, ticks, pellets, water, and hay) to obtain an actual picture of the tularaemia epizootic situation in the Kharkiv, Dnipropetrovsk, and Mykolaiv oblasts. Samples were collected using the flag method (for ticks) and break-back traps (for rodents). Also, hay, water and owl pellets were collected for study. The F. tularensis genetic material in samples was detected using a 16S qPCR. It was found that in Kharkiv oblast, 23% of collected samples were positive for F. tularensis, in Dnipropetrovsk oblast 1.9%, and in Mykolaiv oblast 0.4%. Among the sample types, 34.7% of ticks, 1.8% of rodents, and 36.4% of pellets were positive for F. tularensis. The most frequent carriers of F. tularensis were the D. reticulatus and I. ricinus ticks (74.2% and 29.3%, respectively, of positive results).

Tularaemia is a zoonotic disease caused by the gram-negative bacterium Francisella tularensis, which is endemic to Ukraine. The aim of this work was to provide screening of different field samples (rodent tails, ticks, pellets, water, and hay) to obtain an actual picture of the tularaemia epizootic situation in the Kharkiv, Dnipropetrovsk, and Mykolaiv oblasts.

White sturgeon iridovirus (WSIV) disease is caused by a virus of the eponymous family and is mostly triggered by stressful environmental conditions, i.e. high rearing density, excessive handling, or temporary loss of water. The aim of this study was to develop the most effective diagnostic method for quick and efficient confirmation or exclusion of the presence of WSIV. A total of 42 samples (spleen, gills, intestine, skin, kidney, and brain) were collected from eight sturgeon (Acipenser gueldenstaedtii and A. oxyrinchus) aged ≤5+ farmed or caught between 2010 and 2014 in open waters (Dąbie Lake and Szczecin Lagoon). They were tested for WSIV presence using conventional PCR, qPCR, and in situ hybridisation (ISH). In gross examination, all fish appeared to be healthy. Neither species showed clinical signs typical of WSIV infection. In the majority of cases, fragments of iridoviral DNA were found using molecular methods in the kidneys, and also in the liver, gills, and skin. The detection rate using ISH was 47.37% and most commonly the brain and kidney tissues were positive. The most efficient of the methods used was real-time PCR, with 100% effectiveness in detection of WSIV DNA. The study demonstrates the capabilities for WSIV diagnosis available to sturgeon farmers and water administrators, indicating useful methods of adequate sensitivity as well as organs to sample in order to achieve the highest probability of viral detection.

Introduction White sturgeon iridovirus (WSIV) disease is caused by a virus of the eponymous family and is mostly triggered by stressful environmental conditions, i.e. high rearing density, excessive handling, or temporary loss of water. The aim of this study was to develop the most effective diagnostic method for quick and efficient confirmation or exclusion of the presence of WSIV. Material and Methods A total of 42 samples (spleen, gills, intestine, skin, kidney, and brain) were collected from eight sturgeon (Acipenser gueldenstaedtii and A. oxyrinchus) aged ≤5+ farmed or caught between 2010 and 2014 in open waters (Dąbie Lake and Szczecin Lagoon). They were tested for WSIV presence using conventional PCR, qPCR, and in situ hybridisation (ISH). Results In gross examination, all fish appeared to be healthy. Neither species showed clinical signs typical of WSIV infection. In the majority of cases, fragments of iridoviral DNA were found using molecular methods in the kidneys, and also in the liver, gills, and skin. The detection rate using ISH was 47.37% and most commonly the brain and kidney tissues were positive. The most efficient of the methods used was real-time PCR, with 100% effectiveness in detection of WSIV DNA. Conclusion The study demonstrates the capabilities for WSIV diagnosis available to sturgeon farmers and water administrators, indicating useful methods of adequate sensitivity as well as organs to sample in order to achieve the highest probability of viral detection.

White sturgeon iridovirus (WSIV) disease is caused by a virus of the eponymous family and is mostly triggered by stressful environmental conditions, i.e. high rearing density, excessive handling, or temporary loss of water. The aim of this study was to develop the most effective diagnostic method for quick and efficient confirmation or exclusion of the presence of WSIV.

Q fever (coxiellosis) is an infectious disease of animals and humans, caused by.C. burnetii and widely distributed throughout the world. It is known that people and animals acquire the disease predominantly.via inhalation of infectious aerosols. The possibility of transmission of the pathogen by the alimentary route is still a matter of debate and remains controversial. Therefore the aim of this study was to fill the gaps in knowledge of oral transmission of.C. burnetii by conducting biological tests on the guinea pig model. Guinea pigs, divided into five groups comprising a negative control and four experimental groups, received specified concentrations of.C. burnetii per os. To determine the presence of specific antibodies, blood samples were tested using CFT. Also, internal organs collected during necropsy were screened by a real-time PCR targeting I.1111. Additionally, histopathological evaluation of the tissues was performed. The presence of antibodies and pathogen DNA in caecum was confirmed in one guinea pig from experimental group IV..C. burnetii was also detected in testicular tissue collected from one animal of experimental group II. The presence of pathogen DNA in the testicular tissue indicates that infection spreads haematogenously. In the majority of experimental animals specific antibodies and genetic material of.C. burnetii were not detected. This fact suggests that development of infection depends on many factors, such as animal immune status.

Q fever (coxiellosis) is an infectious disease of animals and humans, caused by.C. burnetii and widely distributed throughout the world. It is known that people and animals acquire the disease predominantly.via inhalation of infectious aerosols. The possibility of transmission of the pathogen by the alimentary route is still a matter of debate and remains controversial. Therefore the aim of this study was to fill the gaps in knowledge of oral transmission of.C. burnetii by conducting biological tests on the guinea pig model.

Introduction: The development of Jembrana disease vaccine is an important effort to prevent losses in the Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine encoding the transmembrane portion of the envelope protein in pEGFP-C1 and test the success of its delivery in culture cells using a chitosan-DNA complex. Material and Methods: Cloning of the DNA vaccine was successfully performed on E. coli DH5α and confirmed by colony PCR, restriction analysis and sequencing. The plasmids were prepared as a chitosan complex using the complex coacervation method and physicochemically characterised using a particle size analyser. A transfection assay was performed in HeLa cells with 4 h exposure, and mRNA expression was assessed at 24 h post transfection. Results: With a 1:2 (wt./wt.) ratio of DNA and chitosan, the complexes have a mean diameter of 236 nm, zeta potential value of + 17.9 mV, and showed no high toxicity potential in the HeLa cells. This complex successfully delivered the DNA into cells, as shown by the presence of a specific RT-PCR product (336 bp). However, the real-time PCR analysis showed that the delivery with chitosan complex resulted in lower target mRNA expression when compared with a commercial transfecting agent. Conclusion: pEGFP-env-tm JDV as a candidate vaccine can be delivered as the chitosan-DNA complex and be expressed at the transcription level in vitro. This initial study will be used for further improvement and evaluation in vivo.

Introduction: The development of Jembrana disease vaccine is an important effort to prevent losses in the Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine encoding the transmembrane portion of the envelope protein in pEGFP-C1 and test the success of its delivery in culture cells using a chitosan-DNA complex.

Serological diagnosis of brucellosis is still a great challenge due to the infeasibility of discriminating infected animals from vaccinated ones, so it is necessary to search for diagnostic biomarkers for differential diagnosis of brucellosis. Cell division cycle 42 (Cdc42) from sheep (Ovis aries) (OaCdc42) was cloned by rapid amplification of cDNA ends (RACE), and then tissue distribution and differential expression levels of OaCdc42 mRNA between infected and vaccinated sheep were analysed by RT-qPCR. The full-length cDNA of OaCdc42 was 1,609 bp containing an open reading frame (ORF) of 576 bp. OaCdc42 mRNAs were detected in the heart, liver, spleen, lung, kidneys, rumen, small intestine, skeletal muscles, and buffy coat, and the highest expression was detected in the small intestine. Compared to the control, the levels of OaCdc42 mRNA from sheep infected with Brucella melitensis or sheep vaccinated with Brucella suis S2 was significantly different (P < 0.01) after 40 and 30 days post-inoculation, respectively. However, the expression of OaCdc42 mRNA was significantly different between vaccinated and infected sheep (P < 0.05 or P < 0.01) on days: 14, 30, and 60 post-inoculation, whereas no significant difference (P > 0.05) was noted 40 days post-inoculation. Moreover, the expression of OaCdc42 from both infected and vaccinated sheep showed irregularity. OaCdc42 is not a good potential diagnostic biomarker for differential diagnosis of brucellosis in sheep.

Serological diagnosis of brucellosis is still a great challenge due to the infeasibility of discriminating infected animals from vaccinated ones, so it is necessary to search for diagnostic biomarkers for differential diagnosis of brucellosis.