Abortion In Sheep Research Articles

Molecular Investigation of Small Ruminant Abortions Using a 10-Plex HRM-qPCR Technique: A Novel Approach in Routine Diagnostics.

The objective of this study was to apply and preliminarily evaluate a High-Resolution Melting (HRM) analysis technique coupled with qPCR, that allows the simultaneous detection of 10 different ruminant abortogenic pathogens, for investigating abortions in sheep and goats throughout Greece. A total of 264 ovine and caprine vaginal swabs were obtained the week following the abortion from aborted females and analyzed using a commercially available kit (ID Gene™ Ruminant Abortion Multiplex HRM, Innovative Diagnostics). Results indicated a high prevalence of Coxiella burnetii and Chlamydophila spp., which were detected in 48.9% and 42.4% of the vaginal swabs, respectively. Results for these most commonly detected pathogens were compared with those of a well-established commercial qPCR kit, with near-perfect agreement. Toxoplasma gondii, Salmonella spp., Brucella spp., Anaplasma phagocytophilum, Campylobacter fetus, and Neospora caninum were also identified, the two latter reported for the first time in the country in small ruminants. Mixed infections occurred in 35.6% of the animals examined. This technique allows for the simultaneous detection of many abortogenic pathogens in an accurate and cost-effective assay. Detection of uncommon or not previously reported pathogens in various cases indicates that their role in ovine and caprine abortions may be underestimated.

Potential role of peste des petits ruminants virus in small ruminant abortions

The aim of the present study was to investigate the frequency, genetic variability, and phylogeny of the peste des petits ruminants virus (PPRV) in ovine and caprine fetuses. During 2014 and 2017, a total of 1054 embryos/fetuses were collected in Turkey. A real-time RT-PCR assay was used for the detection of the PPRV RNA. Genetic characterization and phylogenetic analysis of the PPRV field isolates were conducted by sequencing fusion (F) protein and nucleoprotein (N) gene segments. Samples were also collected from ewes (n = 83) and nanny goats (n = 3) that had aborted and whose embryos/fetuses were found to be PPRV positive. PPRV positive embryos/fetuses were also tested for the presence of Listeria monocytogenes, Campylobacter spp., Coxiella burnetii, Chlamydophila abortus, Brucella spp., akabane virus, aino virus, bluetongue virus, border disease virus, bovine viral diarrhea virus, Cache Valley virus, and Schmallenberg virus. PPRV RNA was detected in 123 (11.7 %) of the 1054 embryos/fetuses, 78 of the 83 (94 %) ewes and 3 (100 %) nanny goats. Border disease virus RNA and Chlamydophila abortus DNA were detected in 7 and 12 PPRV positive sheep fetuses, respectively, while other bacterial and viral agents were not detected. Phylogenetically, the field isolates in this study belong to lineage IV, and compared to other strains of lineage IV considered in this study, they showed 1 and 5 new amino acid substitutions in the F and N gene sequences, respectively. The results of the study suggest that PPRV plays an important role in abortion. Therefore, PPRV needs to be taken into consideration in sheep and goats abortions.

Virulence profiling of Campylobacter spp., C. jejuni and C. fetus subsp. fetus abortions rise in sheep farms in Kashmir, India

Campylobacter spp. are the leading causes of ovine abortions leading to severe economic losses and a source of bacterial food borne illness in humans, posing a major public health concern. This study reports an increase in Brucella negative abortions in sheep farms in Kashmir, India in the last few years. Screening of sheep farms was carried to rule out Campylobacter abortion. Three Government sheep breeding farms in the Kashmir valley and some other private flocks were screened for the presence of C. jejuni and C. fetus subsp. fetus. A total of 217 samples comprising of 200 swabs (rectal and vaginal swabs; 100 each) from clinically healthy animals, ten vaginal swabs from cases of abortion and seven abomasal contents of the aborted fetuses were collected from sheep breeding farm Khimber (District, Srinagar), sheep breeding farm Goabal, the Mountain Research Centre for sheep and goats (MRCSG, SKUAST-K) (District, Ganderbal) and from private sheep breeders were screened. In the present study a total of 15.2% of samples were positive for different Campylobacter spp. by PCR. C. jejuni and C. coli were detected individually or coexisting by PCR in the rectal swabs from all farms, while C. fetus subsp. fetus was detected only in the vaginal swabs from private sheep farms and abortion samples. Additionally, C. jejuni was also isolated from vaginal swabs. A total of 5, 20 and 18 samples were confirmed positive by PCR for C. fetus subsp. fetus, C. jejuni and C. coli, respectively. The Campylobacter isolates obtained in the present study were screened for flaA, cdtB, cadF, wlaN, pldA, virB and dnaJ virulence determinants. However, the isolates harboured flaA, cdtB and cadF virulence determinants only. The recovery of virulent Campylobacter isolates from healthy sheep fecal swabs in the present study may have longer human health implications. The presence of abortive strains of C. jejuni and C fetus subsp. fetus in sheep farms has long term economic implications in the Kashmir valley. This study emphasizes the need for efforts to be taken on farms to prevent animal infections and minimizing human exposure to these pathogens through proper hygiene and production practices as suggested by World Organization for Animal Health (OIE).

Multi-Etiological Abortion due to Campylobacter spp. and Chlamydia abortus in a Sheep

In this case, multi-etiologic abortion due to Campylobacter spp. and Chlamydia abortus was determined in an aborted sheep fetus sent to Konya Veterinary Control Institute (KVKE) from a sheep farm operating in Aksaray province in 2020. The presence of Campylobacter spp. was determined by the bacterial isolation method. C.abortus was identified by qPCR and immunohistochemistry (IHC) methods. This study should be published in order to emphasize that in the fight against sheep abortions, multi-etiologic abortions involving multiple agents should be taken into consideration in addition to a single etiologic agent, and a multidisciplinary approach should be used in the diagnosis to identify the abortion agents.

Biomolecular survey on the main infectious causes of abortion in sheep in the Italian regions of Latium and Tuscany.

Ruminants play an important role in economic sustenance in many developing countries. Abortion is one of the most important causes of economic losses in sheep livestock and, for this reason, it is very important to know, at an early stage, which pathogens caused abortion. The aim of the study is to obtain data about the distribution of abortifacient pathogens in the Italian regions of Latium and Tuscany, the awareness of the distribution of infectious agents causing abortion could allow the development of an appropriate vaccination and prophylaxis plan, to avoid major economic losses. 388 abortions were collected during the 2015-2018 period. Organs, tissues, and swabs were subjected to DNA extraction and then analyzed with commercial q-PCR kits for the detection of the most common abortion pathogens circulating in these geographical areas. The positivity in 148 abortions was 56% for Chlamydia abortus, 14% for Coxiella burnetii, 16% for Salmonella spp, 12% for Toxoplasma gondii, and 2% for Neospora caninum. Interesting results were obtained for cases of abortions with co-infection of abortion pathogens. Diagnosing the cause of abortion remains a multifaceted process that may also include non-infectious factors such as deficiencies and toxicities. Further research is needed also to assess the role of low pathogen concentrations and co-infections in the abortions of sheep.

Sheep abortions associated with Neospora caninum and Toxoplasma gondii infections in multiple flocks from Southern Brazil.

Neosporosis and toxoplasmosis are important parasitic causes of abortions in small ruminants. This study verified the occurrence of these diseases in sheep fetuses from Santa Catarina State, Southern Brazil from 2015 to 2022. Sheep fetuses were necropsied with organ sampling for histopathology, and polymerase chain reaction (PCR) for Neospora caninum and Toxoplasma gondii using the Nc5 and SAG2 targets, respectively, in frozen brain tissue. Microbiological culture and RT-PCR for Pestivirus were conducted to discard other abortion causes. One positive fetus for toxoplasmosis was genotyped using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) with ten genetic markers. Fifty-five sheep fetuses were evaluated, with 10 (18.2%) cases of neosporosis and 7 (12.7%) cases of toxoplasmosis, comprising six and four flocks, respectively. Macroscopically, neosporosis abortions exhibited fetal mummification, maceration, and arthrogryposis. Toxoplasmosis abortions showed fetal mummification and maceration. The neosporosis abortions included lymphoplasmacytic myositis (70%; 7/10) and myocarditis (60%; 6/10), in addition to necrotizing encephalitis and gliosis (50%; 5/10). Toxoplasmosis abortions included lymphoplasmacytic necrotizing encephalitis (71.4%; 5/7), lymphoplasmacytic myositis (42.8%, 3/7), and myocarditis (14.3%; 1/7). Through PCR, N. caninum and T. gondii were detected in 6 (60%) and 5 (71.4%) fetuses, respectively. In one fetus, T. gondii genotyping was conducted, which was characterized as atypical genotype ToxoDB #98. All of the cases were negative for Pestivirus and bacterial agents. This study establishes the occurrence of these diseases as causes of abortions, malformations, mummification, and fetal maceration in sheep, with the characterization of an atypical T. gondii genotype in one of the fetuses.

Review of the epizootic situation on chlamydia in animals and birds in the Russian Federation for the period from 2019 to 2021

Relevance. Animal chlamydia is widespread throughout the world. Chlamydia infection affects animals, birds, fish, reptiles, as well as humans. Causes significant economic damage to livestock farms. Difficulties in diagnosis are associated with pronounced polymorphism of clinical manifestations. According to the order of the Ministry of Agriculture of the Russian Federation of December 19, 2011 No. 476, chlamydia and enzootic abortion of sheep are included in the list of contagious, including especially dangerous, animal diseases, for which restrictive measures (quarantine) can be established. The purpose of our work is to analyze the epizootic situation for chlamydia in animals and birds on the territory of the Russian Federation for the period from 2019 to 2022.Methods. Based on the data obtained from official reports in the 4-vet (annual) form for 2019–2021, an analysis of the epizootic situation and the prevalence of chlamydia in animals and birds in the Russian Federation was carried out.Results. Epizootological monitoring is carried out in all subjects of the Russian Federation. For the period from 2019 to 2021, the State Veterinary Service examined 2 750 726 different biological and pathological materials. There were 11 970 positive results (0.4%) in all federal districts. The highest percentage of positive results was found among pets (dogs and cats). The most informative biological materials for testing for chlamydia are swabs from the mucous membranes and pathological material. The detection rate of positive results using PCR is higher than that of serological methods

Inactivated Flagellin-Containing Vaccine Efficacy against Ovine Enzootic Abortion.

Chlamydia abortus is the etiological agent of abortion and fetal loss in sheep, goats and bovine cattle in many countries. Even though commercially available vaccines can reduce the incidence in sheep, the development of new, safe, and effective vaccines remains high on the agenda. In this study, an evaluation was made of the efficacy of a vaccine candidate, an inactivated antigen based on the extract of outer membrane proteins of a C. abortus strain known as Chlamydia VNITIBP-21, in combination with recombinant flagellin as an adjuvant. Pregnant sheep (n = 43) were divided into three groups: an experimental vaccinated group, a control infected group and a control non-infected group. The sheep were vaccinated twice, with an interval of 3 weeks, then infected with the homologous virulent strain of Chlamydia abortus on pregnancy day 75. The vaccine candidate reduced C. abortus shedding in vaginal swabs considerably, in comparison with the control group. In addition, ewes in the experimental group experienced no abortions, while those in the control group experienced instances of abortion, as well as births of weak and nonviable lambs. The findings show that the vaccine candidate proved itself to be promising in combatting the agent of ovine abortion and fetal loss.

Molecular detection of bacterial zoonotic abortive agents from ruminants in Turkey.

Abortions in cattle and sheep are one of the major causes of economic losses worldwide. Brucella spp. are the most common infectious agent associated with these abortions. However, abortions caused by bacteria such as Listeria spp., Leptospira spp., Campylobacter spp. and Mycoplasma spp. are usually overlooked due to their sporadic nature and their status as non-priority abortion agents. In our study, we investigated the bacteria associated with abortion cases in cattle and sheep using PCR. For this purpose, we collected vaginal swab samples (n: 110) of aborted cattle and sheep, as well as stomach content samples (n: 69) of aborted calves and lambs from various cities in Turkey. The samples were analysed by bacteria-specific PCR to detect Campylobacter fetus, Leptospira spp., Listeria spp., Mycoplasma spp., and Yersinia spp. PCR analyses revealed that the investigated bacterial agents were present in 18.85% and 19.3% of the cattle and sheep samples, respectively, with an overall percentage of 18.99%. While the overall positivity rate for C. fetus, Leptospira spp., and Mycoplasma spp. was 2.79%, 10.06%, and 4.47%, respectively, the positivity rate for co-infection with Leptospira spp. and C. fetus was 1.68%. All samples were found to be negative for Yersinia spp. and Listeria spp. The high C. fetus positivity rate detected in sheep and in the stomach contents was statistically significant (p < 0.05). However, the difference in positivity rates between the cities, hosts, co-infections and causative agents was statistically insignificant (p > 0.05). This study provides preliminary data on the significant involvement of C. fetus, Leptospira spp. and Mycoplasma spp. in cattle and sheep abortions in Turkey indicating that they should not be overlooked in diagnosis. In addition, further research is needed to investigate the zoonotic potential of these pathogens for public health in Turkey.

Development of a Novel Real-Time PCR-Based Test Panel System for Detection of Infectious Abortion in Domestic Sheep

Development of a Novel Real-Time PCR-Based Test Panel System for Detection of Infectious Abortion in Domestic Sheep

Distribution of Species and Biotypes of Brucella Isolates Obtained from Sheep and Cattle Abortions

Distribution of Species and Biotypes of Brucella Isolates Obtained from Sheep and Cattle Abortions

Multietiological abortion induced by Brucella melitensis and Chlamydophila abortus in a sheep fetus

In this case, multietiological abortion due to Brucella melitensis (B. melitensis) and Chlamydophila abortus (C. abortus) agents was defined in a sheep abortion. An aborted sheep fetus submitted to Konya Veterinary Control Institute (KVCI) from a sheep farm in Nigde province in Brucella spp. presence was determined by bacterial isolation method and confirmed by slide agglutination test with monospecific A and M sera. C. abortus was detected by real time PCR and immunohistochemistry (IHC) methods. In histopathological examinations, in the lung; hyperemia, hemorrhage and bronchopneumonia, In the heart; in the placenta with degeneration and necrosis of heart muscle cells; hyperemia, hemorrhage, edema, necrosis and mononuclear cell infiltrations were observed. In this study, it was emphasized that in the fight against sheep abortions, multi-etiological abortions involving more than one factor should be taken into consideration, as well as an etiological agent, and it would be appropriate to use multidisciplinary diagnostic methods to determine the abortion factors, thus contributing to the prevention of abortions, which is still an important problem in sheep breeding.

Molecular Survey of Toxoplasma gondii Infection in Aborted Fetuses of Sheep in the Iğdır Province of Türkiye

Toxoplasma gondii, an obligatory intracellular protozoan parasite, can infect a wide range of warm-blooded animals, including livestock species. T. gondii is a zoonotic protozoan parasite that affects both humans and other warm-blooded animals. The aim of this study was to detect T. gondii by using PCR in the brain tissues of 60 aborted sheep fetuses from the Iğdır Province in Türkiye. For this purpose, 60 brain tissue samples of sheep were collected within the lambing seasons of 2023 in Iğdır, Türkiye. The DNA extraction was performed using the PureLink™ Genomic DNA Mini Kit from brain samples. The PCR was performed with the appropriate primers from the obtained DNA samples. T. gondii was found in the brain (16.6%) samples of aborted sheep fetuses. According to the present study, T. gondii infection can be one of the causes of fetus abortion of sheep in Iğdır province, Türkiye. This result emphasizes the need for vigilance and preventive measures in managing this potential public and animal health concerns.

Molecular investigation of Coxiella burnetii in aborted fetus of small ruminants in southeast Iran.

The domestic animal, known as a main reservoir of Coxiella burnetii, is susceptible to the occurrence of coxiellosis, which can lead to abortions in domestic animals, causing significant economic damage and posing risks to human health. Therefore, the purpose of this study is to investigate C. burnetii as the causative agent of Q fever in abortion samples of small ruminants in southeastern Iran. This study was conducted between 2020 and 2021 in Zarand city, located in Kerman province (southeast Iran). In this study, 50 abomasum swab samples of aborted sheep and goat fetuses were collected and analyzed using molecular methods to identify C. burnetii. The results revealed that 26% (n: 13) of the collected abortion samples were infected with C. burnetii. Among the positive samples, two (50%) belonged to goat abortion samples while 11 (23.9%) belonged to sheep abortion samples. This study demonstrates that C. burnetii is one of the causes of abortion in small ruminants in southeastern Iran. It is recommended to pay more attention to C. burnetii in domestic animals due to its significant economic impact on livestock and its potential implication for human health in Iran.

Campylobacter species abortion in sheep.

This focus article was prepared by Jasmine Smith of the APHA Small Ruminant Species Expert Group. Data extraction and analysis was done by Anna Brzozowska of the APHA Surveillance Intelligence Unit and Leanne Dempsey from APHA Thirsk.

An Early Treatment With BKI-1748 Exhibits Full Protection Against Abortion and Congenital Infection in Sheep Experimentally Infected With Toxoplasma gondii.

Congenital toxoplasmosis in humans and in other mammalian species, such as small ruminants, is a well-known cause of abortion and fetal malformations. The calcium-dependent protein kinase 1 (CDPK1) inhibitor BKI-1748 has shown a promising safety profile for its use in humans and a good efficacy against Toxoplasma gondii infection in vitro and in mouse models. Ten doses of BKI-1748 given every other day orally in sheep at 15 mg/kg did not show systemic or pregnancy-related toxicity. In sheep experimentally infected at 90 days of pregnancy with 1000 TgShSp1 oocysts, the BKI-1748 treatment administered from 48 hours after infection led to complete protection against abortion and congenital infection. In addition, compared to infected/untreated sheep, treated sheep showed a drastically lower rectal temperature increase and none showed IgG seroconversion throughout the study. In conclusion, BKI-1748 treatment in pregnant sheep starting at 48 hours after infection was fully effective against congenital toxoplasmosis.

Border disease virus and chlamydophila abortus co-infection in aborted sheep foetuses

Abortion in sheep and goats is one of the most important reproductive problems that affect small ruminants breeding. Non-infectious factors and infectious agents such as bacterial, viral, mycotic and parasitic infectious agents can cause abortion in sheep and goats. A high rate of abortion (43.75%) was observed in two sheep flocks in Niğde Province in Turkey during the lambing season in 2016. To determine the reason of abortion, aborted sheep foetuses (n = 3) and EDTA whole blood samples (n = 3) from mother of the foetuses were collected from two sheep flocks. Aborted foetuses and buffy coat cells of the EDTA whole blood samples were used for total nucleic acid extraction. Extracted nucleic acids were analysed for akabane virus (AKAV), bluetongue virus (BTV), peste des petits ruminants virus (PPRV), pestiviruses, Schmallenberg virus (SBV), Brucella spp., Chlamydophila abortus (C. abortus), Coxiella burnetii and Listeria monocytogenes. Border disease virus (BDV) RNA and C. abortus DNA were detected in three aborted sheep foetuses whereas other investigated infectious agents were not detected. Additionally, EDTA whole blood samples from mother of the foetuses were also found BDV positive. To the best of my knowledge, this is the first report on BDV and C. abortus co-infection in aborted sheep foetuses.

An In house ELISA using recombinant GRA1 and BAG1 proteins as antigen successfully detected Toxoplasma gondii infection in a flock of sheep suffering from abortions

Toxoplasma gondii is one of the main pathogens causing abortion in sheep. In this study, an in house ELISA using recombinant GRA1 and BAG1 proteins as antigen was used to detect T. gondii infection in a flock of sheep suffering from abortions. Serum samples collected from sheep (n = 23) the day after abortion were initially analyzed with a commercial ELISA using recombinant P30 (SAG1) protein as antigen and then by the in house ELISA using recombinant GRA1 and BAG1 proteins as antigen. Commercial ELISA detected anti-T. gondii IgG antibodies in 20 samples whereas in house ELISA detected anti-T. gondii IgG antibodies in 19 samples. The results of the remaining three serum samples that were seronegative by commercial ELISA were also seronegative by in house ELISA. According to the results of commercial ELISA and in house ELISA, the seroprevalence of ovine toxoplasmosis was 86.9% and 82.6%, respectively. When we accept the validated commercial ELISA as a reference method, the sensitivity and specificity of in house ELISA can be considered as 95% and 100%. These preliminary results show that in house ELISA using recombinant GRA1 and BAG1 proteins as antigen can detect ovine toxoplasmosis successfully.

Türkiye’de kızıl tilki beyin (Vulpes vulpes) örneğinde Neospora caninum’un ilk moleküler tespiti

The red fox is the wild carnivore with the widest distribution in the world. Thus, this animal acts as intermediate and final host for many parasite species. Neospora caninum is one of the most important protozoan agents causing abortion in cattle, sheep and goats in the world. The final hosts of N. caninum are domestic dogs and wild canids such as wolves and coyotes, while its intermediate hosts are domestic ruminants and many warm-blooded animals, including red foxes. The aim of this study was to research N. caninum in brain samples of three red foxes obtained from wildlife in Türkiye by using PCR. At the end of the study N. caninum DNA was detected in one of three brain samples. To the best of our knowledge, with this study, N. caninum was detected for the first time in a red fox brain sample in Türkiye.

Molecular detection and phylogenic characterization of Neospora caninum in naturally infected sheep in Alborz and Qazvin provinces, the north of the central region of Iran.

Neospora caninum is a protozoan coccidian parasite that can act as a cause of abortion in sheep. The aim of this study was to investigate the presence of this parasitic agent and its role in causing abortion in sheep of Iran. Between June 2019 and February 2022, 100 samples [brain (n = 39), placenta (n = 8), embryonic membrane (n = 7), cotyledon (n = 7), umbilical cord (n = 2), homogenate mixture of tissues (heart, liver, spleen and digestive track) (n = 37)] that were collected following the necropsies of 39 aborted ovine fetuses from different parts of the Alborz and Qazvin provinces, the north of the central region of Iran were employed for DNA extraction. Nc-5 was selected as the target gene sequence for amplification of DNA by using four pairs of primers in two semi-nested PCR. Samples considered positive for the presence of the NC-5 gene were examined to further confirm the presence of the ITS1 gene. Sequence of NC-5 gene was detected from the 27 tissue samples of 23 aborted ovine fetuses. The ITS1 gene sequence was detected in all of the 27 tissue samples that were positive for the NC-5 gene analysis. Brain tissue was the most studied tissue, and the highest number of positive cases was observed in this tissue. The present study updated the situation of ovine neosporosis in the central region of Iran and confirmed the presence of the N. caninum among sheep flocks' abortion.