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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.

Abortion is one of the major threats to the livestock industry, and it also poses significant threats to public health since some of the abortifacient agents are considered zoonotic. Chlamydia abortus (C. abortus), Coxiella burnetii (C. burnetii), Listeria monocytogenes (L. monocytogenes), and Cache Valley virus (CVV) are recognized as important zoonotic and abortifacient agents of reproductive failure in small ruminants. This study determined the prevalence of these agents in ovine and caprine foetuses in Türkiye. A total of 1 226 foetuses were collected from the sheep (n = 1 144) and goats (n = 82) from different flocks between 2012 and 2017. Molecular detection methods were used to detect C. abortus, C. burnetii, and L. monocytogenes DNA and CVV RNA in aborted foetuses. In this study, C. abortus was the most prevalent abortifacient agent among the investigated ovine (264/1144) and caprine (12/82) foetuses, followed by C. burnetii with a frequency of 2.8% (32/1144) and 8.5% (7/82) in ovine and caprine foetuses, respectively. L. monocytogenes DNA was detected in 28 (2.4%) and 2 (2.4%) of the ovine and caprine foetuses, respectively. However, CVV RNA was not detected. Although the predominant mixed infection was C. abortus and C. burnetii, mixed infection of C. abortus and L. monocytogenes, and C. burnetii and L. monocytogenes were also found. The information presented in this study contributes to the understanding of the roles of C. abortus, C. burnetii, L. monocytogenes, and CVV in abortions in small ruminants, and could be beneficial for developing more effective control strategies.

Bovine viral diarrhea virus (BVDV) is the most prevalent pathogen in cattle and causes significant economic losses due to its severe clinical manifestations. It belongs to the family Flaviviridae and is distributed in species A, B, and H within the genus Pestivirus. The objective of this study was to detect and characterize BVDV using molecular techniques (RT-PCR) in semen and aborted fetuses samples that were sent to the CEDIVEP (Veterinary Diagnostic Center of Paraguay) laboratory. Seventy-three samples of semen from bulls were analyzed, and 54.7% of the samples were positive for Pestivirus A. The presence of Pestivirus A and H was detected in 2/8 spontaneously aborted fetuses. The genotypes of four individual samples of type A and four samples of type H organs were confirmed by partial sequencing of the 5-UTR region. The presence of BVDV was confirmed by molecular techniques for the first time in our country through its detection in different types of samples, as well as the presence of two genotypes. This suggests that the circulation of this virus can cause significant losses in cattle production in Paraguay.

Background Equine herpesvirus type 1 (EHV-1) is commonly associated with horse abortion. Currently, there are no reported cases of abortion resulting from EHV-1 infection in donkeys. Results This was the first survey-based study of Chinese donkeys. The presence of EHV-1 was identified by PCR. This survey was conducted in Chabuchar County, North Xinjiang, China, in 2020. A donkey EHV-1 strain (Chabuchar/2020) was successfully isolated in MDBK cells. Seventy-two of 100 donkey sera were able to neutralize the isolated EHV-1. Moreover, the ORF33 sequence of the donkey-origin EHV-1 Chabuchar/2020 strain showed high levels of similarity in both its nucleotide (99.7‒100%) and amino acid (99.5‒100%) sequences, with those of horse EHV-1 strains. EHV-1 Chabuchar/2020 showed significant consistency and was classified within cluster 1 of horse EHV-1 strains. Further, analysis of the expected ORF30 nucleotide sequence revealed that donkey EHV-1 strains contained guanine at position 2254, resulting in a change to aspartic acid at position 752 of the viral DNA polymerase. Therefore, these strains were classified as horse neuropathogenic strains. Lastly, a phylogenetic tree was constructed using the partial ORF68 nucleotide sequences, showing that the identified donkey EHV-1 strain and the EHV-1 strain found in aborted Yili horses in China comprised a novel independent VIII group. Conclusion This study showed the first isolation and identification of EHV-1 as an etiological agent of abortions in donkeys. Further analysis of the ORF33, ORF30, and ORF68 sequences indicated that the donkey EHV-1 contained the neuropathogenic genotype of strains in the VIII group. It is thus important to be aware of EHV-1 infection in the donkey population, even though the virus has only been identified in donkey abortions in China.

Bovine herpesvirus 4 (BoHV-4) is increasingly believed to be responsible for several disorders of the bovine reproductive tract. The first characterization of BoHV-4 in Argentina was from samples from an aborted fetus. Argentinean isolates are highly diverse and are phylogenetically grouped in three genotypes. In this study, we describe the isolation of BoHV-4 from a bovine fetus with a gestational age of 8 months and without macroscopic lesions. Genetic analyses revealed that the isolated strain belongs to genotype 2. This is the first report on the presence of infectious BoHV-4 in tissues from an aborted bovine fetus.

Porcine circovirus type 2 (PCV2) is a ubiquitous pathogen, and co-infections with the emerging PCV3 are increasingly reported. Both PCV2 and PCV3 have been implicated in reproductive failure, yet the diagnostic criteria for PCV3 remain under development. While fetal or neonatal antibody detection is a recognized indicator of transplacental infection in multiple species, PCV2 appears to be an exception due to the possible transfer of maternal antibodies. This study evaluated IgG, IgA, and IgM antibodies in the heart, kidney, lung, and spleen of aborted fetuses from sows co-infected with PCV2 and PCV3. PCR analysis revealed that all aborted fetuses were positive for both PCV2 and PCV3, with PCV3 Ct values being generally lower than those of PCV2, although this difference was not statistically significant. Antibody profiling showed a higher prevalence of anti-PCV3 IgM and IgA compared to anti-PCV2 IgM and IgA, particularly in the heart, kidney, and lung, while IgG responses against both viruses were similar. These findings suggest that the detection of anti-PCV3 antibodies in fetal tissues may provide supportive evidence of PCV2 and PCV3 infection and the possible involvement of these viruses in reproductive failure; however, further studies are needed to establish causation definitively.

Objective Abortion emerges as a primary concern for small ruminant breeders, leading to substantial economic losses for livestock farmers. The most important factor contributing to abortion in sheep and goats is the presence of infectious agents. The bluetongue virus (BTV), border disease virus (BDV), peste des petits ruminants virus (PPRV), Akabane virus (AKAV) and pox virus are notable viruses responsible for inducing abortion in small ruminants globally. Methods This study focused on the prevalence of viral agents in aborted foetuses and adult sheep and goats with a recent abortion history in East Azerbaijan province, Northwest Iran. For this purpose, a comprehensive evaluation was performed in the nine cities of East Azerbaijan province, Iran. A total of 62 aborted foetuses and 373 blood samples were collected from 43 sheep and goat flocks. The conventional and nested PCR methods were employed for BTV, BDV, PPRV, pox virus and AKAV detection after extracting viral DNA/RNA and cDNA synthesis from the buffy coat (adults) and foetal spleen tissue samples. The formalin‐fixed tissue samples from the aborted foetuses were examined histopathologically. Results Molecular findings revealed that viral infection was present in 7.77% (29 out of 373) of the blood samples and 35.48% (22 out of 62) of the aborted foetuses. Among the blood samples, 1.34%, 2.41% and 4.29% were positive for BTV, BDV and PPR, respectively. No pox virus or AKAV samples tested positive in the adults. In the samples from aborted foetuses, the positivity rates were 12.90%, 25.80%, 9.67%, 1.61% and 0% for BTV, BDV, PPRV, pox virus and AKAV, respectively. Histopathological studies revealed extensive inflammatory reactions associated with severe hyperaemia and haemorrhagic lesions in the tissue sections, particularly in the brain, heart, intestine and spleen, which were more notable in the PCR‐positive samples for BTV. Conclusion In conclusion, the detection of viral agents in both aborted foetuses and blood samples indicates that viral infections play notable roles in the abortion of sheep and goats in East Azerbaijan. Therefore, effective management and vaccination strategies are crucial for preventing and controlling the disease in this region. Evaluation of viral causes of abortion in small ruminants in East Azerbaijan province, Northwest Iran. Step 1: Sample collection. a. A total of 373 blood samples were collected from adult sheep and goats with a recent abortion history. b. A total of 62 aborted foetuses were collected after a systematic necropsy. Step 2: The blood samples (buffy coats) were evaluated using the conventional PCR. Among these, 1.34%, 2.41% and 4.29% were positive for BTV, BDV and PPR, respectively. Step 3: The spleen of the aborted foetuses were evaluated using the conventional PCR. Among these, the positivity rates were 12.90%, 25.80%, 9.67%, 1.61% and 0% for BTV, BDV, PPRV, pox virus and AKAV, respectively. Moreover, the tissues were examined using pathological studies, which revealed extensive vascular and inflammatory reactions in the intestine (A), spleen (B), liver (C), heart (D) and brain (E).

Background Equid alphaherpesvirus 1 (EHV-1) is one of the main infectious causative agents of abortion in mares and can also be associated with stillbirth, neonatal foal death, rhinopneumonitis in young horses and a neurological disorder called equine herpesvirus myeloencephalopathy (EHM). The neuropathogenicity of the virus was shown to be significantly higher in EHV-1 strains that carry a single nucleotide point (SNP) mutation in the ORF30, which encodes a catalytic subunit of viral DNA polymerase (ORF30 D 752 ). Another gene, ORF68 is frequently used for phylogenetic analysis of EHV-1. Methods 27 EHV-1 strains isolated from aborted equine fetuses in Poland, collected between 1993 and 2017, were subjected to PCR targeting the open reading frames (ORFs) 30 and 68 of the EHV-1 genome. PCR products obtained were sequenced and SNPs were analyzed and compared to sequences available in GenBank. Results None of the analyzed sequences belonged to the ORF30 D 752 neuropathogenic genotype: all EHV-1 belonged to the non-neuropathogenic variant N 752 . On the basis of ORF68 sequences, the majority of EHV-1 sequences (76.9%) cannot be assigned to any of the known groups; only six sequences (23.1%) clustered within groups II and IV. Conclusions EHV-1 strains obtained from abortion cases belong to the non-neuropathogenic genotype. Many EHV-1 ORF68 sequences have similar SNPs to those already described in Poland, but a clear geographical distribution was not observed. A single particular ORF68 sequence type was observed in strains isolated from 2001 onwards.

Background To date, four human bocaviruses (HBoV) have been described. The most closely related viruses (bovine and canine parvoviruses) are associated with miscarriage in their hosts. The objective of this retrospective study was to determine the frequency of HBoV DNA in miscarriage. Study Design Tissue samples from 172 patients, in which miscarriage occurred, were included and tested with a published qPCR protocol. Positive PCRs were mutually confirmed by sequencing. Results 43 patients (25%) were positive for HBoV DNA. Of those, the majority of HBoV‐positive samples were tissues from miscarriage (placenta: 6; aborted tissue products of conception: 37 specimens). The samples were not paired; either placental or aborted tissue was available. Conclusions The results show that, as long as no animal model is available, the role of HBoV in the occurrence of miscarriage requires additional prospective studies in order to investigate its significance and causal involvements of this pathogen.

Background. Parvovirus B19 infection during pregnancy can lead to nonimmune fetal hydrops, miscarriage, and intrauterine fetal death (IUFD). Some studies have suggested that parvovirus B19 infection may surprisingly often result in nonhydropic fetal death during the third trimester, in the absence of maternal serological evidence of acute infection. This study was conducted to investigate the prevalence of parvovirus B19 DNA among fetuses from miscarriages and IUFDs. Methods. We retrospectively studied 535 unborn fetuses, including 120 fetuses from miscarriages and 169 from IUFDs. The control fetuses were 246 fetuses from induced abortions. All fetuses were autopsied from July 1992 through December 1995 and from January 2003 through December 2005 in Helsinki, Finland. The period included a major epidemic of parvovirus B19 infection in 1993. Formalin-fixed, paraffin-embedded fetal tissues were studied with use of a highly sensitive and specific PCR that was capable of detecting all 3 parvovirus B19 genotypes and by histologic examination. In addition, maternal parvovirus B19 serological status was determined. Results. Parvovirus B19 DNA was detected in 5 fetuses with gestational ages of 14, 22, 23, 30, and 39 weeks; these included fetuses from 4 (2.4%) of the 169 IUFDs and 1 (0.8%) of the 120 miscarriages. During the epidemic year 1993, the prevalence of parvovirus B19 DNA–positive fetal deaths was 6 times the prevalence during nonepidemic years. All 5 mothers of the parvovirus B19 DNA–positive fetuses had serological signs of acute parvovirus B19 infection close to the time of fetal death. The only nonhydropic fetus was full-term. Conclusions. Our findings indicate that the prevalence of parvovirus B19 infection among fetuses from IUFDs is low. In particular, our findings did not verify the claimed high prevalence of third-trimester nonhydropic IUFDs associated with parvovirus B19.