Canine Leishmaniosis in Greece: An Updated Countrywide Serological Study and Associated Risk Factors

Canine leishmaniosis (Leishmania infantum) is a zoonotic disease that affects dogs worldwide. Greece is enzootic for this disease, and updated data for its current distribution are of major importance. The aim of this cross-sectional serological study was primarily to update the current knowledge of Leishmania infantum seropositivity status within the asymptomatic Greek canine population and, furthermore, to assess the possible climatological and other risk factors. In total, sera of 1265 asymptomatic dogs were collected from all prefectures of the country. A questionnaire that included all individual dog information was completed for all animals. The Speed Leish K® canine Leishmania antibody test kit (BVT Groupe Virbac, France) was employed. Potential risk factors were evaluated utilizing logistic regression models. Overall, 13.8% (n = 175) of the sampled dogs were seropositive to Leishmania infantum originating from all geographical departments of the country, whereas most prefectures had at least one seropositive animal. Outdoor living, high mean humidity, low mean wind speed and high total annual rainfall were found to increase the seropositivity status against the parasite. Conclusively, Leishmania infantum remains a common parasite challenge in the asymptomatic canine population of Greece, and therefore, its early diagnosis and effective prevention are significant in the country.

Seroprevalence and Risk Factors Associated with Brucella Infection in Camels in the Puntland State of Somalia

Brucellosis is an important zoonotic disease caused by members of the genus Brucella. Camel brucellosis has been reported in almost all camel-rearing countries in Africa and Asia. A cross-sectional study was conducted between February 2020 and February 2021 in Galkayo, Garowe, and Bosaso districts in the Puntland State of Somalia to investigate the seroprevalence and risk factors of brucellosis in camels. A competitive enzyme-linked immunosorbent assay (c-ELISA) was used to detect anti-Brucella antibodies, while a structured questionnaire was used to collect epidemiological data. A total of 441 camel sera were screened against Brucella antibodies. Thirty-one (7%; 95% CI: 4.8–9.8%) samples were positive, and thirteen (54.2%; 95% CI: 32.8–74.4%) out of the twenty-four farms sampled had at least one seropositive animal. Galkayo district had the highest number of Brucella-seropositive camels (10.3%), followed by Bosaso district (8.6%), while Garowe district had the lowest number of seropositive camels (1.4%). The binary logistic regression model revealed that camels in Galkayo district (p = 0.015; OR: 9.428; 95% CI: 1.539–57.755), camels from large herd sizes of >50 animals (p = 0.019; OR: 5.822; 95% CI: 1.336–25.371), and those in contact with small ruminants (p = 0.011; OR: 10.941; 95% CI: 1.728–69.285) were significantly associated with seropositive cases of camel brucellosis in the Puntland State of Somalia. The present study shows that Brucella infections in camels are prevalent in the three districts covered by the study. This poses a public health risk, because milk from these camels is used for human consumption. Studies focusing on the isolation of Brucella strains in camels and investigating brucellosis in ruminants and humans are recommended in the study area. Validation of serological tests—including c-ELISA—for Brucella antibody detection in camels is also needed.

Prevalence of Toxoplasma gondii, Hepatitis E Virus and Salmonella antibodies in meat juice samples from pigs at slaughter in Switzerland

Toxoplasma gondii , hepatitis E virus (HEV) and Salmonella are zoonotic foodborne pathogens that may be transmitted to humans through the consumption of raw or undercooked pork. The aim of this study was to determine the seroprevalence of anti- Toxoplasma gondii , anti-HEV and anti- Salmonella antibodies from healthy pigs at slaughter in Switzerland. In the period of August to September 2020 diaphragm muscle of Swiss fattening pigs was collected in three Swiss abattoirs from a total of 188 farms. Two randomly chosen pig carcasses per farm were selected. On the basis of the slaughter data, the production system and the canton of origin were noted, comparing indoor (n=120) and free-range farming (n=68), and regional allocation. The meat juice of these samples was analyzed for pathogen-specific antibodies using commercial enzyme-linked immunosorbent assay (ELISA) kits. The seroprevalences were 1.3% for T. gondii , 71.8% for the HEV and 5.3% for Salmonella , respectively. Comparing the origins, the results of many cantons weren't meaningful due to the low number of samples. No regional accumulations were found for T. gondii and the HEV. The results showed that 2.1% of the farms had least one T. gondii seropositive animal, 80.3% had at least one HEV seropositive animal, and 8.5% had at least one Salmonella seropositive animal, respectively. The seropositivity of T. gondi i was higher in free-range pigs than in indoor pigs, whereas anti- Salmonella antibodies were more common in pigs from indoor farming than in outdoor pigs. The seroprevalence of anti HEV-Abs was similar in free-range and indoor farming pigs. Compared to studies from 2012 the seroprevalence of T. gondii has decreased whereas the seroprevalence of the HEV has increased and is highly prevalent among fattening pigs in Switzerland. The low seroprevalence of Salmonella has remained stable in recent years.

A Freedom of Coxiella burnetii Infection Survey in European Bison (Bison bonasus) in Poland

Q fever is an important zoonosis caused by the intracellular Gram-negative bacteria Coxiella burnetii. The source of infection are numerous species of mammals, birds, reptiles and amphibians, as well as ticks. The disease is widespread throughout Europe, but the role of wildlife in its epidemiology is poorly understood. The European bison (Bison bonasus) population has been growing European-wide quite dynamically over the last few years. The aim of this study was to determine whether C. burnetii infection occurs in European bison and whether it can be considered an important bacterial reservoir in the natural environment. Five hundred and twenty three samples of European bison sera originating from 14 (out of the 26 existing) Polish populations were examined for the presence of specific antibodies using an ID Screen Q Fever Indirect Multi-species ELISA test. Only one (0.19%) serum sample was positive in ELISA, and two other samples were doubtful. The only seropositive animal found in this study was a free-living bull. It suggests possible transmission from domestic cattle by sharing pastures. The transmission of C. burnetii into the European bison was rather accidental in the country and its role as an important wild reservoir is unlikely. Since no tests are available for wildlife ruminants there is a need for the adaptation of the available tests.

Risk Factors Associated with Seroprevalence of Chlamydia abortus in Sheep Farms in Ceará, Brazil

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortions and birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, and the microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it a public health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibody levels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil (Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited. Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using an Enzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordance with the manufacturer's instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks. Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds had at least one seropositive animal. The number of seropositive individuals was significantly different between adults and ewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modeling identified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR = 2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus antibodies in sheep in the State of Ceara emphasizes the importance of testing and tracking the disease spread among herds; these results were similar to studies in other areas of Brazil. Adult sheep that spend more time on the property may have a higher exposure risk because of increased reproductive activity. Misinformation and technical limitations can influence the proper handling of animals avoiding contagion through the correct use of techniques and recommendations. Disease transmission occurs through the digestive tract and between mother and fetus. Therefore, seropositive (infected) sheep may be related to the breeding system practices, such as allowing contact between sheep and other species on the property (goats, cattle, and horses) during breeding. Acquiring animals from external sources without sufficient health information can increase the transmission risk. Contaminated pastures, water, food, and air also increase transmission risk. The lack of technical and practical knowledge regarding disease prevention and control also contributes to disease transmission, resulting in reproductive losses due to high abortion rates. Chlamydia abortus has zoonotic potential and may infect humans without proper safety information. Therefore, future epidemiological studies are required for a better understanding of the primary risk factors for disease occurrence and spread among herds in the region. Chlamydia abortus infection is present in sheep in Ceará, Brazil. Chlamydiosis information programs should be adopted, sanitary measures implemented, and the epidemiological surveillance of sheep herds strengthened.

Spatial Distribution of Serologically Reactive Sheep to Leptospira spp. in the Northeast Region of Brazil

Background: Considering the importance of leptospirosis in sheep farming and public health and the significance of identifying which serogroups circulate in sheep within each region, the objective of this study was to determine the seroprevalence and spatial distribution of the most frequent serogroups causing infection by Leptospira sp. in ovine herds in the Northeast region of Brazil.Materials, Methods & Results: Blood samples were collected from 4197 sheep from 229 herds in 7 Northeastern States. Sera were analyzed via microscopic agglutination test (MAT). The frequency of seroreactive sheep for Leptospira sp. was 14.06%. The states of Alagoas, Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe, located in the Caatinga biome, had the highest frequencies of serologically reactive sheep, and Maranhão, in the Cerrado biome, had the lowest frequency. The most frequent serogroups were Autumnalis (19.49%), Australis (15.76%) and Serjoe (14.41%). In the states of Ceará, Paraíba, Rio Grande do Norte, and Sergipe, 100% of their municipalities had at least one seroreactive animal. The highest frequencies of seropositive animals were found in the municipalities of União (50%), Passagem (49.06%), Canindé (48.89%), Igaci (28.95%), Gararu (31.2%), Pirapemas (17.5%), and Angicos (16%) located in the states of Piauí, Paraíba, Ceará, Alagoas, Sergipe, Maranhão and Rio Grande do Norte, respectively.Discussion: The animal-level prevalence (14.06%) obtained in the present study is significant, especially considering the rustic nature of the species and the adverse conditions of the region for the infectious agent. In semi-arid conditions, it has been suggested that perhaps sheep do not seroconvert detectable titers on MAT with a cut-off point of 1:100. It is important to highlight that theovine population in the Northeast region of Brazil is composed of mixed animals, which have been considered more resistant to infection by Leptospira spp. Also, environmental factors hostile to the survival of the infectious agent in the studied region should be taken into consideration, since they may have influenced the seropositive animal-level prevalence. A noteworthy variation was observed in agglutinin titers, which ranged from 100 to 1,600, where 80.2% of the positive samples had titers ≤ 200. It is important to highlight that more elevated titers (≥ 400) were obtained in all seven states, which may suggest an acute infection caused by a non-adapted serovar, indicating that preventive and control measures focused on possible infection sources for sheep should be adopted. Although some states showed the same serogroups as the most frequent, a variety of serogroups was observed in municipalities, which may indicate different sources of infection, whether interspecies, intraspecies, or via alternative routes of transmission in semi-arid conditions, such as venereal. This indicates that even though sheep are more resistant to infection, they become exposed due to the environment or management conditions. As such, identification, isolation, and treatment of the affected animals are alternative measures recommended for prevention and control of leptospirosis in sheep in the semi-arid region. It is evident that despite the lack of rain observed in the last decade in the Northeast region of Brazil, which prevented the formation of favorable environments for the presence of Leptospira, the infectious agent remains among the sheep, as well as other production and wild animals in the region. Some factors may be contributing to this scenario, such as the fact that sheep farming in the region is characterized mainly by subsistence systems, where veterinary assistance and adequate sanitary management are absent, thus increasing the possibility of contact with Leptospira.Keywords: small ruminants, semi-arid, zoonosis.

Distribution of Toxoplasma gondii Tissue Cysts in Shoulder Muscles of Naturally Infected Goats and Lambs

ABSTRACT Toxoplasmosis has been recognized as a major public health problem worldwide. The consumption of uncooked or undercooked meat infected with Toxoplasma gondii tissue cysts is one of the main means of transmission of this parasite. Although sheep, goats, and pigs are commonly infected with T. gondii, little information is available on the distribution of T. gondii tissue cysts in naturally infected meat. In this study, we investigated the distribution of viable T. gondii tissue cysts in shoulder muscles of naturally infected lambs and goats. Hearts and shoulders of 46 lambs and 39 goats from a local grocery store were tested for T. gondii infection. Animals were evaluated for the presence of anti–T. gondii antibodies in heart blood and clots by the modified agglutination test. Fourteen of the 85 animals (seven lambs and seven goats) were seropositive. Six to 12 samples weighing 5, 10, and 50 g were obtained from shoulder muscles of each seropositive animal and used for bioassay in mice. The distribution of viable T. gondii differed according to the size of the sample analyzed, but in general larger sample sizes resulted in higher isolation rates (P &lt; 0.05). Results of the study revealed an uneven distribution of T. gondii in muscle samples of lambs and goats and that T. gondii can be transmitted by consumption of very small servings (5 and 10 g) of meat when it is consumed raw or is undercooked. HIGHLIGHTS

Cross-sectional survey for toxoplasmosis in free-range chickens (Gallus gallus domesticus) from the Atlantic Forest area in Alagoas state, Northeastern Brazil

Toxoplasmosis is a parasitic disease, which is prevalent across the world and has a strong environmental component in its transmission chain. The present study determined the seroprevalence and factors related to Toxoplasma gondii infection among free-range chickens from an Atlantic Forest area in Northeastern Brazil. A total of 550 free-range chickens were tested for T. gondii antibodies usingthe indirect fluorescent antibody test (IFAT, cut-off point 1:16), and 269 (48.9%; 95% CI = 44.7-53.1%) were positive. Among the 61 properties 57 (93.4%) had at least one seropositive animal. The robust Poisson regression model showed that the variables/categories associated with the seroprevalence of T. gondii were: region of origin of the animals/urban area (prevalence ratio [PR] = 2.346; P<0.001), management system/use of cages (PR = 1.591; P = 0.019), presence of rodents/yes (PR = 1.295; P = 0.035), and the type of food/use of food scraps (PR = 1.603; P = 0.009). The high prevalence found suggest that this scenario demands careful management, mainly regarding the use of scraps of undercooked or raw food, adequate environmental hygiene and frequent rodentcontrol.

Positive association between Brucella spp seroprevalences in livestock and humans from a cross-sectional study in Garissa and Tana River Counties, Kenya

BackgroundBrucella spp. is a zoonotic bacterial agent of high public health and socio-economic importance. It infects many species of animals including wildlife, and people may get exposed through direct contact with an infected animal or consumption of raw or undercooked animal products. We implemented a linked livestock-human cross-sectional study to determine seroprevalences and risk factors of Brucella spp. in livestock and humans. We also estimated intra-cluster correlation coefficients (ICCs) for these observations at the household and village levels.MethodologyThe study was implemented in Garissa County (specifically Ijara and Sangailu areas) and Tana River (Bura and Hola) counties. A household was the unit of analysis and the sample size was derived using the standard procedures. Serum samples were obtained from selected livestock and people from randomly selected households. Humans were sampled in both counties while livestock could be sampled only in Tana River County. Samples obtained were screened for anti-Brucella IgG antibodies using ELISA kits. Data were analyzed using generalized linear mixed effects logistic regression models with the household (herd) and village being used as random effects.ResultsThe overall seroprevalences were 3.47% (95% confidence interval [CI]: 2.72 – 4.36%) and 35.81% (95% CI: 32.87 – 38.84) in livestock and humans, respectively. In livestock, older animals and those sampled in Hola had significantly higher seroprevalences that younger ones or those sampled in Bura. Herd and village random effects were significant and ICC estimates associated with these variables were 0.40 (95% CI: 0.22 – 0.60) and 0.24 (95% CI: 0.08 – 0.52), respectively. For human data, older people, males, and people who lived in pastoral areas had significantly higher Brucella spp. seroprevalences than younger ones, females or those who lived in irrigated or riverine areas. People from households that had at least one seropositive animal were 3.35 (95% CI: 1.51 – 7.41) times more likely to be seropositive compared to those that did not. Human exposures significantly clustered at the household level; the ICC estimate obtained was 0.21 (95% CI: 0.06 – 0.52).ConclusionThe presence of a seropositive animal in a household significantly increased the risk of exposure in people in that household. Brucella spp. exposures in both livestock and humans clustered significantly at the household level. This suggests that risk-based surveillance measures, guided by locations of primary cases reported, either in humans or livestock, can be used to detect Brucella spp. infections livestock or humans, respectively.Author summaryBrucellosis is an important zoonotic disease that primarily affects livestock and wildlife. In humans, the disease is characterized by prolonged fever, body aches, joint pains and weakness while in livestock, the disease causes abortions and infertility. We carried out a study in northeastern Kenya (Garissa and Tana River Counties) to identify factors that affect the distribution of the disease in people and livestock. Livestock and people from randomly selected households were recruited and serum samples obtained for screening using anti-Brucella IgG ELISA kits to determine their Brucella spp. exposure. Data obtained were analyzed using mixed effects logistic regression models. Results obtained show that human and animal Brucella spp. seroprevalences cluster at the household level. The odds of exposure in humans were at least three times higher in households that had at least one seropositive animal compared to those that did not. These results can be used to design risk-based surveillance systems where each Brucella infection identified in livestock or humans could signal potential locations of other secondary infections.

C. burnetii Shedding Study In Domestic Animals in Georgia

ObjectiveQ fever is poorly understood in Georgia and its prevalence is largely underestimated in both humans and animals.One of the main goal of the project was shedding study in domestic animals – isolation of C. burnetii from suspected seropositive animal blood, milk samples.IntroductionQ fever is a zoonotic bacterial disease resulting from infection by Coxiella burnetii. Domestic ruminants (cattle, sheep, and goats) are considered the main reservoir for the pathogen, which can also infect humans. Q fever is poorly understood in Georgia and its prevalence is largely underestimated in both humans and animals.In Georgia Q fever laboratory diagnostic was started and implemented at the Laboratory of the Ministry of Georgia (LMA) within GG20 ,,Prevalence, Epidemiological Surveillance, and Laboratory Analysis of Coxiella burnetii in Georgia’’.MethodsLMA conducted Coxiella burnetii shedding evaluation in three specific farms from Kvemo Kartli (Tsalka, Dmanisi) and Mtskheta-Mtianeti (Dusheti). Seropositive cattle and small ruminants were sampled per week. Sampling lasted 7 weeks and totally 581 samples samples (blood, milk and swab) were tested. Testing were conducted in a BSL3 laboratory under BSL3 working conditions. ACCM medium was used (2XACCm-2 acidified Citrate Cysteine Medium PH-4.75G N NaOH). The samples were incubated at 37°C using CO2.ResultsAs a result of the study, one culture was bacteriologically isolated from seropositive cattle milk sample ( the sample was taken on the third week of the study in Beshtasheni farm, Tslka, Kvemo Kartli) and confirmed by Molecular biology (PCR).ConclusionsThe study confirmed Q fever existence in Georgia. Traditionally considered an obligate intracellular agent, the requirement to be grown in tissue culture cells, embryonated eggs, or animal hosts has made it difficult to isolate C. Burnetii strains. Within the study one culture was isolated from the seropositive animal milk sample that was collected in the third week of the study. shedding of Coxiella burnetii in milk by infected cows appeared to be the most frequent positive sample for the bacterium.