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Objectives: The study aimed to detect Neospora caninum by nested PCR (nPCR) in aborted fetuses of cattle, sheep, and goats in Bangladesh. Materials and Methods: The head portion of each aborted fetus (111) was dissected at each sampling site and transferred to the laboratory in an ice box. Data on risk factors associated with N. caninum infection were simultaneously collected. Deoxyribonucleic acid was extracted from brain tissue to perform nPCR targeting the internal transcribed spacer 1 (ITS1) ribosomal DNA (rDNA) gene of N. caninum and sequencing was performed from the representative positive samples. Results: By nPCR, N. caninum was found in 16.0% of aborted fetuses of cattle, followed by sheep (14.81%) and goats (11.78%). The highest prevalence was found in aborted fetuses of animals during the second trimester (27.78%) of pregnancy aged 2 to 4 years (18.75%). Obtained sequences showed they were completely matched with N. caninum ITS1 rDNA gene deposited in GenBank. Univariate analysis demonstrated that pregnancy stages (trimesters), abortion history of the animals, and access to dogs in animal farms were significantly (p ≤ 0.05) correlated with N. caninum infection. Conclusion: This study represents the first investigation into the molecular detection, phylogenetic characterization, and analysis of risk factors associated with N. caninum in livestock in Bangladesh. According to the research findings, N. caninum infection may have a role in abortion cases and the ensuing financial losses in the nation’s livestock industry.

Porcine circovirus 3 (PCV-3) has been widely detected in healthy and diseased pigs; among different pathologic conditions, the strongest evidence of association comes from reproductive disease cases. However, simple viral detection does not imply the causality of the clinical conditions. Detection of PCV-3 within lesions may provide stronger evidence of causality. Thus, this study aimed to assess the frequency of PCV-3 detection in tissues from fetuses/stillborn piglets in cases of reproductive problems in domestic swine, as well as the histopathologic assessment of fetal tissues. Fetuses or stillborn piglets from 53 cases of reproductive failure were collected and analyzed by PCV-3 qPCR. The presence of porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus 2 (PCV-2), and porcine parvovirus 1 (PPV1) was also checked. PCV-3 qPCR positive samples with a high viral load were tested by PCV-3 in situ hybridization (ISH), sequenced, and phylogenetically analyzed. PCV-3 DNA was detected in 18/53 (33.9%) reproductive failure cases and in 16 of them PCV-3 was the only pathogen found. PCV-2 DNA was found in 5/53 (9.4%), PRRSV RNA in 4/53 (7.5%) and PPV1 was not detected. Four out of the six PCV-3 qPCR-positive cases with Ct value <30 were positive when tested by ISH. In these samples, PCV-3 was detected within mild histopathologic lesions, such as arteritis and periarteritis in multiple tissues. The present work emphasizes the need to include PCV-3 as a potential causative agent of reproductive failure in swine.

We aimed to identify DNA in the brain samples of aborted fetuses of cattle, goats, and sheep in Mazandaran, northern Iran, using PCR. In total, 133 aborted fetuses (51 sheep, 78 cattle, and 4 goats) were randomly collected from different stages of gestation in various regions of Mazandaran, Iran, from Mar 2016 to May 2017. The DNA was extracted from all the brain samples using phenol chloroform isoamyl alcohol instructions. The gene was used for the detection of DNA by nested-PCR assay. The detection of DNA was confirmed by the observation of a 227 bp band in 24 samples of 133 aborted fetuses (18.1%). The highest prevalence rate of was detected in the cattle (20.5%) followed by the sheep (15.6%); however, no positive cases were reported in the goats. The highest and lowest prevalence rates of the infection were reported as 23.8% and 8.6% in Qaemshahr, and Behshahr, respectively. The prevalence rate of infection (32%) in the early gestational period was higher than those in the middle (15%) and late (3.8%) gestational periods. The obtained data of the present study indicated that infection may partly be responsible for abortion and economic loss in livestock farming in Mazandaran Province.

Congenital toxoplasmosis can cause severe consequences in the fetus, such as spontaneous abortion which is affected by parasite strain. Also, recent studies revealed the high genetic diversity of Toxoplasma gondii . This study aims to investigate the serological status of T. gondii in pregnant women, multilocus genotyping in aborted fetuses’ tissue, and archived formalin‐fixed paraffin-embedded placenta. This study was performed on 100 pregnant women with spontaneous abortion and their aborted fetuses, and 250 of the archived placentae in Iran. The blood and tissue were examined for seroprevalence and genotype determination of T. gondii using ELISA and multilocus nested-PCR–RFLP, respectively. Anti- T. gondii IgG and IgM were detected in 68 samples (68%) and 1 (1%) out of 100 serums. Toxoplasma DNA was identified in 1 (1%) aborted fetuses’ tissue and 32 (12.8%) placenta samples. Overall, ten positive DNA samples were successfully genotyped, and five genotypes were recognized (ToxoDB#1, #2, #10, #27, and #48). The obtained results indicated congenital toxoplasmosis is a severe risk in this region. As type I is highly pathogen and can lead to severe complications, the prevention of the infection should be considered in seronegative pregnant women.

Background: We aimed to detect Toxoplasma gondii in ovine aborted fetuses and evaluate its genetic variations in the southwest of Iran. Methods: This cross-sectional study was performed on 100 aborted ovine fetuses collected from the different region of Kohgiluyeh and Boyer-Ahmad Province, Iran, in lambing season during 2017 and 2018. DNA was extracted from the brain samples of all of the aborted fetuses and PCR amplified, targeting a 529 bp repetitive element gene of T. gondii. Moreover, to find out the heterogeneity of the positive samples, PCR-DNA amplification of the two main genetic markers, B1 and GRA6, of T. gondii were performed. Nucleotide sequencing and phylogenetic analysis were performed, using the BLAST program and MEGA-X software. Results: The 529 bp gene of T. gondii was detected in 2 out of 100 (2%) of the ovine aborted samples. The sequences analysis of GRA6 and B1 genes revealed that both isolates from the aborted fetuses of sheep belonged to type I of T. gondii. Intra-divergence was more seen in GRA6 gene whereas less divergence was observed in B1 gene. Conclusion: Congenital infection with Type I of T. gondii during the neonatal period is associated with abortion in ovine. Evaluation of more aborted samples from broader geographical areas is needed to elucidate the molecular epidemiology and also the genotypes of T. gondii associated with abortion.

Background: Toxoplasma gondii is a zoonotic obligatory intracellular protozoan parasite that infects a wide range of warm-blooded species. This study aimed to obtain further information on the role of T. gondii infection in ruminant abortion (sheep, goats and cattle) using bioassay and PCR methods in Mazandaran province, northern Iran. Methods: Overall, 104 aborted fetuses (52 bovine, 48 ovine, 4 caprine) were collected at different stages of gestation during the lambing seasons in various parts of Mazandaran Province from Mar 2016 to May 2017. Brains of 104 aborted fetuses were bioassayed in female BALB/c mice. DNA was extracted from all brain samples using phenol-chloroformisoamyl Alcohol instructions. RE gene was used for detection all of T. gondii DNA by conventional PCR assay. Results: The results of the bioassayed samples were negative because no tachyzoites or cyst were observed in the peritoneal and brain specimens of the mice. The detection of T. gondii DNA was confirmed by observation of a 529 bp band in 15 out of 104 fetuses (14.4%). The highest prevalence rate of T. gondii detected from sheep (16.6%) followed by cattle (13.4%) and goats (0%). The highest prevalence of the infection was observed in east area, while the lowest prevalence of the infection was observed in west area. Conclusion: T. gondii infection may partly be responsible for abortion and economic losses in livestock husbandry in this region. Therefore, further additional researches such as genotyping T. gondii and designing control strategies for improving management in livestock flocks are necessary.

Porcine circovirus 3 (PCV-3) has been associated with several pig diseases. Despite the pathogenicity of this virus has not been completely clarified, reproductive disorders are consistently associated with its infection. The aim of the present work was to analyze the presence of PCV-3 DNA in tissues from pig fetuses from different gestational timepoints. The fetuses were obtained either from farms with no reproductive problems (NRP, n = 249; all of them from the last third of gestation) or from a slaughterhouse (S, n = 51; 49 of the second-third of gestation and 2 from the third one). Tissues collected included brain, heart, lung, kidney, and/or spleen. Overall, the frequency of detection of PCV-3 was significantly higher in fetuses from the last third of the gestation (69/251, 27.5%) when compared to those from the second-third (5/49, 10.2%), although the viral loads were not significantly different. Moreover, the frequency of detection in NRP fetuses (69/249, 27.7%) was significantly higher than in S ones (5/51, 9.8%). Furthermore, PCV-3 DNA was detected in all tissue types analyzed. In conclusion, the present study demonstrates a higher frequency of PCV-3 DNA detection in fetuses from late periods of the gestation and highlights wide organ distributions of the virus in pig fetuses.

Background Infectious abortion in ruminants is a problem in animal husbandry worldwide. It is important to obtain a diagnosis, to make sure that proper control measures can be instituted, but most abortion cases remain without an etiologic diagnosis. This report describes the presence of Arcobacter species and several neglected opportunistic abortifacient agents in ruminant abortion cases showing or not co-infections among at least one of the major recognized protozoal, fungal, bacterial and viral abortifacient agents. Results A total of 67 fetuses (55 cattle and 12 goats) and just one placenta (cattle) were considered. Among the most common abortive agents, Neospora caninum (19,4%), followed by Chlamydophila abortus (4,5%), Listeria monocytogenes 1/2a (2,98%), Bovine Viral Diarrhea Virus type 1b (2,98%), Bovine herpesvirus 4 (2,98%), and Aspergillus spp. (2,98%) were detected. The isolated neglected opportunistic bacteria include Escherichia coli , Acinetobacter lwoffii , Staphylococcus spp., Streptococcus spp., Streptococcus uberis , Streptococcus suis , Trueperella pyogenes, Mannheimia haemolytica , Bacillus cereus and Nocardia spp. Other bacterial species, not associated with abortion by literature, but described as causes of diseases occurring sporadically both in humans and animals, were also detected. Three Arcobacter strains, namely two A. skirrowii and one A. cryaerophilus , were isolated from 3 bovine aborted fetuses, and A. butzleri was isolated from the placenta. Conclusions A not negligible isolation of Arcobacter species and other neglected abortifacient agents has to be mentioned, with prevalences that seem to be emerging and replacing or co-placing the major infectious players in bovine and caprine reproductive failure due to abortion disease, even if further studies investigating the aetiological power and transmission routes are needed in order to define the role of these microrganisms in ruminant abortion.

The only genus of the Francisellaceae family known to contain species pathogenic to mammals is Francisella, for which reported cases in the Southern Hemisphere have been limited to Australia. We describe severe necrotizing and inflammatory lesions and intralesional immunohistochemical identification of Francisella sp. lipopolysaccharide among aborted ovine fetuses in Uruguay.

This present study was carried out to use conventional and real-time PCR for detection and segregation of Brucella abortus and Brucella melitensis in aborted bovine, ovine, caprine, buffalo and camel fetuses. All samples were collected and immediately transferred to laboratory, genomic DNA was extracted and the conventional and real-time PCR by specific primers for Brucella abortus and Brucella melitensis was performed. A TaqMan analysis and single-step PCR was carried out in total 3710 DNA of abomasal contents of aborted fetuses. In total, 281/892 (31.5%) bovine, 224/810 (27.65%) ovine, 219/786 (27.86%) caprine, 199/604 (32.94%) buffalo and 201/618 (32.52%) camel fetus samples gave positive results for Brucella species by conventional PCR. Moreover, 45/281 and 231/281, 169/224 and 49/224, 194/219 and 22/219, 57/199 and 137/199 and finally 51/201 and 143/201 specimens were positive for B. melitensis and B. abortus in aborted bovine, ovine, caprine, buffalo and camel fetuses by real-time PCR, respectively. The sensitivity and specificity of real-time PCR obtained 100% and 100%. Statistical analysis showed significant differences (p<0.01) between B. abortus and B. melitensis that were detected in abomasal contents of aborted bovine, ovine, caprine, buffalo and camelid fetuses and between presences of Brucella spp. in bovine with caprine, buffalo and camel aborted fetuses (p<0.05). The CT values obtained from real-time PCR had significant differences between aborted bovine, ovine, caprine, buffalo and camel fetuses for presence of B. abortus and B. melitensis. Results showed that the real-time PCR is considerably faster than current standard methods for isolation and segregation of Brucella spp. [PUBLICATION ABSTRACT]