Detection and phylogenetic characterization of astroviruses in insectivorous bats from Central‐Southern Italy

In recent years, bats have been found to harbour many viruses, raising several questions about their role as reservoirs and potential disseminators of zoonotic viruses. We investigated the presence of six virus families in bats in three regions of Central‐Southern Italy. Astroviruses were identified in seven of 13 bat species. Sequence analysis revealed marked genetic heterogeneity among the astroviruses identified, with nucleotide identity ranging between 60.26% and 87.62%. Astrovirus diversity was not associated with the bat species, the geographic areas or the bat colony, suggesting the circulation of several astrovirus strains in Italian ecosystems. Genetic diversification and interspecies transmission appear common in bat astroviruses and could provide, potentially, the bases for transmission to humans and other mammals. Yet overemphasizing this risk might have detrimental consequences for bat conservation and preservation of the important ecosystem services bats provide.     

Survey on vertical infection of bovine viral diarrhea virus from fetal bovine sera in the field

Bovine viral diarrhea virus (BVDV) isolation and antibody survey were performed using 2,758 fetal bovine sera (FBS) collected from slaughterhouses in New Zealand, Australia and the Dominican Republic, and then sent to Japan to manufacture commercial serum for cell culture use. FBS in the Dominican Republic were pooled for each several individuals, and those collected in other countries were separated according to each individual and subjected to the tests. BVDV was isolated from 25 (0.91%) FBS, and the BVDV antibody was detected in 44 (1.60%) FBS. The survey on 139 sets of paired sera of a dam and her fetus revealed that neither the BVDV antibody nor BVDV was detected in all FBS from BVDV antibody-positive dams.     

Comparison of carcass and meat characteristics of Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis)

This study was conducted to determine and compare the live traits, carcass yield and the physical, chemical, processing and sensory properties of meat from 20 Brahman grade cattle (crossbred cattle) and crossbred water buffalo (Philippine carabao × Murrah, Bulgarian or Indian) with an average age and weight of 29 months and 434 kg, respectively. The animals were fed a similar diet for 180 days before slaughter. A survey was performed to establish consumer meat preference between the two species. Live weights before slaughter of the two species were found to be different (P < 0.05) with the crossbred water buffalo showing higher bodyweight. However, the crossbred water buffalo had a lower (P < 0.05) dressing yield based on hot carcass weight and chilled carcass weight. The lower dressing yield of crossbred water buffalo can be attributed to its higher (P < 0.05) weights of edible and non‐edible slaughter by‐products. The chilled carcass yield of the forequarters and hindquarters from crossbred cattle and crossbred water buffalo were comparable when expressed as percent of live weight and chilled carcass weight. The estimated lean yield was higher in crossbred cattle than crossbred water buffalo. Crude protein, ash, fat, cholesterol, myofibrillar, sarcoplasmic and insoluble protein contents of the beef and carabao meat were all similar. Water holding capacity, pH, muscle fiber diameter, tenderness, firmness and marbling score in carabeef were all comparable to the beef. Redness was found to be higher (P < 0.05) in carabeef than the beef as measured with a color meter. The consumer meat preference survey showed that 55.9% selected the beef while 44.1% preferred carabeef. The color and amount of fat on the outside of the beef sample were primary in buyer selection.     

Endemic infections of bovine viral diarrhea virus genotypes 1b and 2a isolated from cattle in Japan between 2014 and 2020

Bovine viral diarrhea virus (BVDV) is a causative agent of bovine viral diarrhea. In Japan, a previous study reported that subgenotype 1b viruses were predominant until 2014. Because there is little information regarding the recent epidemiological status of BVDV circulating in Japan, we performed genetic characterization of 909 BVDV isolates obtained between 2014 and 2020. We found that 657 and 252 isolates were classified as BVDV-1 and BVDV-2, respectively, and that they were further subdivided into 1a (35 isolates, 3.9%), 1b (588, 64.7%), 1c (34, 3.7%), and 2a (252, 27.7%). Phylogenetic analysis using entire E2 coding sequence revealed that a major domestic cluster in Japan among BVDV-1b and 2a viruses were unchanged from a previous study conducted from 2006 to 2014. These results provide updated information concerning the epidemic strain of BVDV in Japan, which would be helpful for appropriate vaccine selection.     

Genetic and antigenic analysis of Chlamydia pecorum strains isolated from calves with diarrhea

Chlamydia pecorum (designated 22–58) was isolated in 2010 in HmLu-1 cells from the jejunum of a calf which died of necrotizing enterocolitis in Yamaguchi Prefecture, Japan. Immunohistochemical staining identified C. pecorum positive reactions in the jejunal villi. C. pecorum, designated 24–100, was isolated from the feces of a calf with diarrhea in another farm in Yamaguchi Prefecture in 2012. A significant increase in neutralizing antibody titers against C. pecorum was confirmed in paired sera. Nucleotide sequence identities of omp1 genes of the 2 isolates were 100%. The isolates were genetically and antigenically more closely related to C. pecorum Bo/Yokohama strain isolated from cattle with enteritis in Japan than to the other prototype strains, Bo/Maeda isolated from cattle with pneumonia and Ov/IPA isolated from sheep with polyarthritis. These results indicate that C. pecorum strains similar to 22–58 and 24–100 might be endemic in Yamaguchi Prefecture and cause enteric disease in cattle.     

Prevalence and molecular analysis of Sarcocystis infections in cattle in Northwest Iran and the first global report of S. gigantea in cattle

Cattle are intermediate host for several species of Sarcocystis, including S. cruzi, S. hirsuta, and S. hominis with high prevalence worldwide. The present study aimed to determine the prevalence of Sarcocystis infection, species identification, and phylogenetic analysis of the parasite in cattle in Northwest Iran. The samples of diaphragm and esophagus from 290 cattle were collected from slaughterhouses in Northwest Iran and subjected to macroscopic, microscopic, and histopathology examinations, PCR-RFLP, sequencing and phylogenetic analyses. Tissue cysts of Sarcocystis spp. were detected in 92% of cattle by digestion and microscopic tests. Based on the PCR–RFLP and specific PCR, 87.9% and 1.03% of isolates were identified as S. cruzi, and S. hominis, respectively. Macrocyst was seen in a single sample that was identified as S. gigantea. The haplotype network exhibited the extension of the various haplotypes of S. cruzi between neighboring provinces in Northwest Iran. Heterogeneity analysis of S. cruzi 18S-rRNA sequences indicated genetic diversity among S. cruzi isolates (Haplotype diversity: 0.733-0.854) consisting 16 haplotypes; however, the nucleotide differences showed low diversity (0.01481 to 0.03351). Pair wise sequence distance matrix amongst S. cruzi sequences indicated an intra-species divergence of 0%-7.8% and identity of 92.6%-100%. Sarcocystis infection is highly prevalent in cattle in Northwest Iran, with the predominance of S. cruzi, and genetic variants of this species are unequivocally distributing in Northwest provinces. First global detection of S. gigantea in cattle reflects new insights of transmission dynamic and biology of this parasite in Iran.     

Comparison of feed intake, digestibility and fattening performance of Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis)

Twenty growing crossbred cattle and crossbred water buffalo (carabao) with an average age of 22 (18–24 months) months were equally distributed into two treatment groups according to species. The animals were fed with the same ration made up of corn silage (50%) + wet brewer's spent grain (30%) + concentrate mixture (20%), and their fattening performance was monitored. The digestibilities of the different nutrients were likewise determined. The economics of raising the animals under intensive production system was calculated. Species differences did not influence total dry matter intake of the animals, when expressed as percentage of the bodyweight and per metabolic body size. There were no significant differences in digestion coefficients of the different nutrients, except for crude protein in crossbred water buffalo and crossbred cattle, although the digestibility of dry matter, organic matter and nitrogen free extract tended to be high in the former than in the latter. Likewise, average daily gain (ADG) was similar, although crossbred water buffalo had numerically higher ADG (828.6 vs 785.5 g) than crossbred cattle during the 6 months feeding. During the first 3 months of feeding (1–90 days), the ADG of crossbred water buffalo was 1066.1 g compared to 940.1 g for crossbred cattle. From 91 to 180 days, the crossbred cattle had slightly higher ADG (630.1 vs 591.1 g) but also the difference was not significant. The return above feed cost was comparable for crossbred cattle and crossbred water buffalo during the first 90 days of feeding. However, extending the feeding period from 91 to 180 days , income over feed cost was higher (P < 0.05) for crossbred cattle by PhP 5.3/kg gain than crossbred water buffalo. Results showed that crossbred water buffalo could attain similar growth rate with that of crossbred cattle under intensive system, when fed with high quality feed materials.     

Comparison of carcass and meat characteristics of Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis) fed on high roughage diet

The objective of this study was to compare the carcass and meat quality in crossbred cattle and crossbred water buffalo at the same younger age and fed with high roughage in the Philippines. Ten crossbred cattle and 10 crossbred water buffalo, with an average age of 22 months (18–24 months) were used in this experiment. The animals were fed a similar diet, which consisted of 85% Napier or Para grass and 15% concentrate mixture on a dry matter basis, for 180 days before slaughter. Slaughter weight of the crossbred water buffalo was higher (P < 0.05) than that of the cattle, although the dressing percentage was significantly (P < 0.01) lower for the former than for the latter. The estimated lean yield was higher (P < 0.05) in crossbred cattle than crossbred water buffalo. The muscle fiber diameter of loin in buffalo meat was smaller (P < 0.05) than that in cattle. In regard to carcass and meat quality, the crossbred water buffalo was comparable or slightly superior to the crossbred cattle in tenderness, sensory score of color (P < 0.01) and flavor (P < 0.05). These results clearly indicate that at a younger age and even under high roughage‐based fattening rations, crossbred water buffalo are able to produce good quality meat, which will be by no means inferior to that of crossbred cattle in the Philippines.     

Comparison of fattening performance in Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis) fed on high roughage diet

The objective of this study was to compare the fattening performance in Brahman grade cattle (crossbred cattle) and crossbred water buffalo at the same young age and fed with high roughage based fattening rations in the Philippines. Ten crossbred cattle and 10 crossbred water buffalo, aged between 18 and 24 months old were used in this experiment. The animals were fed diets consisting of 85% Napier or Para grass and 15% concentrate mixture (CM) on a dry matter (DM) basis. The grass, total DM intake and bodyweight gain were significantly (P < 0.01) higher for the crossbred water buffalo than for the cattle. There was no species significant difference in the digestion coefficient and feed conversion rate between the crossbred cattle and water buffalo. The return over feed cost for fattening was significantly (P < 0.05) higher in the crossbred water buffalo than in the cattle. These results clearly indicate that under high roughage‐based fattening rations, young crossbred water buffalo are better able to utilize the roughage and they perform better in terms of feed intake and live weight gains than the crossbred cattle in the Philippines.     

Molecular characterization of bovine foamy virus and its association with bovine leukemia virus infection in Vietnamese cattle

The detection of bovine foamy virus (BFV) in Vietnamese cattle was performed using conventional PCR targeting pol and gag genes. Out of 243 tested samples, ten (4.1%) and eight (3.3%) samples were positive for BFV gag and pol DNA, respectively. The prevalence of bovine leukemia virus (BLV) estimated by detection of proviral DNA using nested PCR targeting env gene was 26.7% (65/243). The results of nucleotide sequence alignment and the phylogenetic analysis suggested that Vietnamese BFV strains showed high homology to isolates belonging to either European or non-European clades. There was no significant correlation between BLV and BFV. This study provides information regarding BFV infection and confirms the existence of two BFV clades among Vietnamese cattle for the first time.     

1株牛病毒性腹泻病毒河北株的分离鉴定与遗传演化分析

从河北省保定市某养殖场采集疑似患病毒性腹泻/黏膜病(BVD)的犊牛粪便6份,进行牛病毒性腹泻病毒(BVDV)的分离培养,根据GenBank上登录的BVDV5'-UTR片段设计引物,利用RT-PCR技术对目的片段进行扩增,扩增得到的目的基因片段送至公司测序,测序结果进行同源性分析和系统进化树构建.结果:分离出1株致细胞病...     

Molecular phylogenetic analysis of bovine papular stomatitis viruses detected in Saga,Japan

In this study, we performed a molecular phylogenetic analysis of six bovine papular stomatitis virus (BPSV) field strains detected from Japanese beef calves kept on a farm in Saga prefecture, a southwest part of Japan, from 2017 to 2020. The phylogenetic analysis based on a partial B2L gene (554-nt) showed that these field strains were divided into two lineages, a lineage (A-lineage) constructed by a Saga strain and strains obtained from various regions of Japan and the world, and other lineage (B-lineage) constructed by five Saga strains and strains obtained from France, USA and Iwate prefecture (a north part of Japan). Furthermore, a Saga field strain named BPSV_SAGAbv2 and strains obtained from USA and Iwate prefecture belonged to a sub-lineage blanched from B-lineage. This is the first report elucidating molecular epidemiological characters of field BPSVs obtained from Saga prefecture. The existence of the multiple lineages was thought to be related to a history of calf introduction from various regions of Japan into Saga prefecture.     

Horizontal transmission and phylogenetic analysis of bovine leukemia virus in two districts of Miyazaki,Japan

Horizontal transmission is recognized as a major infection route for bovine leukemia virus (BLV), and cattle with high viral loads are considered to be a major infectious source in a herd. However, a correlation between viral loads and the risk of infection has been insufficient to use as a foundation for BLV control strategies. In this report, we examined the epidemiology of BLV infection and the infectious source in a local area. In 2013–2014, BLV infection was investigated in 1,823 cattle from 117 farms in two adjacent districts, Miyazaki, Japan. Seropositive samples for BLV were detected with 88 cattle and in 14 farms. Phylogenetic analysis revealed that 94% of the isolates clustered into genotype I and the remaining isolate into genotype III. Among genotype I, genetically distinct strains were spread at each farm, and cattle infected with less than 3 copies/100 cells did not transmit BLV to other cattle for more than thirty months. This is the first report of concrete data of viral load in relation to viral horizontal transmission under the field condition. The data facilitate farmers and veterinarians understanding the status of BLV infected cattle. This research contributes to BLV infection control and the development of effective BLV eradication programs.     

牛新孢子虫病和弓形虫病的流行病学调查

为了调查我国牛新孢子虫和弓形虫的感染情况，应用新孢子虫酶联免疫吸附试验（ELISA）和弓形虫间接血凝试验（IAT）分别检测了来自国内10个省（市、自治区）的262份乳牛、10份肉牛和40份水牛血清。结果显示，乳牛新孢子虫的抗体阳性率为17．2％，弓形虫的抗体阳性率为2．3％，没有检测到既有新抱子虫抗体又有弓形虫抗体的乳牛血清。各牛场所检乳牛血清的新孢子虫抗体阳性率在0～34．4％之间。在水牛和肉牛血清中未检测到新孢子虫抗体和弓形虫抗体。流产乳牛血清的新孢子虫抗体阳性率为20．2％，未流产乳牛为16．1％，其中血清抗体阳性乳牛主要在妊娠中晚期流产。各个年龄段乳牛血清的抗体阳性率差异不显著（P〉0．05）。不同妊娠胎次的乳牛血清抗体阳性率差异不显著（P〉0．05）。确认新孢子虫病在国内大部分牛场存在。     

Epidemiology and genetic characterization of BVDV,BHV-1, BHV-4, BHV-5 and Brucella spp. infections in cattle in Turkey

The aim of the study was to determine the epidemiological data of bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV-1), bovine herpesvirus-4 (BHV-4), bovine herpesvirus-5 (BHV-5) and Brucella–associated cattle that were previously reported to have abortion and infertility problems in Ankara, Corum, Kirikkale and Yozgat provinces, Turkey. Whole blood and sera samples were obtained from 656 cattle, and antibodies against Brucella spp. were detected in 45 (6.86%) and 41 (6.25%) animals by Rose Bengal plate and serum tube agglutination tests, respectively. The seropositivity rates against BVDV, BHV-1 and BHV-4 were 70.89%, 41.3% and 28.78%, respectively. RT-PCR and PCR were performed to detect RNA and DNA viruses in blood samples, respectively. The BVDV 5′-untranslated region and BHV-1 gB gene detected in this study were phylogenetically analyzed. The BVDV strains analyzed in this study were closely related to those previously reported from Turkey. The nucleotide sequence from the BHV-1 strain detected in this study is the first nucleotide sequence of BHV-1 circulating in this area of Turkey deposited in the GenBank. The presence of Brucella spp. and prevalence of BHV-1, BHV-4 and BVDV in cattle should be further investigated throughout these regions.     

Evaluation of antimicrobial use in dairy cattle,beef cattle and broilers in Japan using dosage-based indicators

The sales amount of antimicrobials intended for use in dairy cattle, beef cattle and broilers from 2008 to 2019 was evaluated for each antimicrobial class and administration route using dosage-based indicators. Our results revealed that the antimicrobial sales amount sold for use in dairy cattle in 2019 in terms of total weight of active ingredient, the number of defined daily doses (DDDs) (theoretical amount of biomass subjected to antimicrobial treatment in a year) and the number of treatment days (TDs) (theoretical number of days of treatment that an animal is subjected to in a year) calculated using Japanese DDD values (DDDjp values) was 36,751 kg, 8,261,848,000 kg·days and 15.5 days, respectively. Likewise, the antimicrobial sales amount sold for use in beef cattle and broilers in 2019 in terms of these metrics was 33,403 kg, 3,928,248,000 kg·days and 3.61 days, and 69,773 kg, 2,947,848,000 kg·days and 10.66 days, respectively. There was a considerable difference between the number of DDDs calculated using DDDjp values and that calculated using European DDD values (DDDvet values) for antimicrobial products sold for use in dairy and beef cattle. Our results also revealed that the sales amount of some antimicrobials, such as cephalosporins and quinolones represented larger proportions when calculated using dosage-based indicators than when calculated using the weight of active ingredient. Considering that Japanese veterinarians and farmers are more likely to conform to the Japanese dosage recommendations rather than the European ones, these results indicate the need for using dosage-based metrics, in particular metrics based on Japanese dosages rather than European dosages.     

Three cases of idiopathic eosinophilic enteritis with chronic obstinate diarrhea in Japanese Black fattening cattle

Eosinophilic enteritis (EOE) is a type of inflammatory bowel disease and is characterized clinically by chronic obstinate diarrhea. Three Japanese Black (JB) fattening cattle (2 males and 1 female) on different cattle farms presented with chronic episodic diarrhea without fever or dehydration. Soft reddish spherical carneous tissues (1−3 cm) were occasionally excreted within the diarrheic feces. Administration of antibiotics, antidiarrheal drugs and vermicides had no therapeutic effect, but dexamethasone improved the fecal characteristics. The symptoms persisted until the animals were slaughtered at 27–30 months of age. Histopathological examination of the intestines revealed marked eosinophilic infiltration in the lamina propria and submucosa. From these findings, we diagnosed these cattle as the first cases of EOE in JB cattle.     

Genetic characterization of bovine viral diarrhea virus strains in Beijing,China and innate immune responses of peripheral blood mononuclear cells in persistently infected dairy cattle

To acquire epidemiological data on the bovine viral diarrhea virus (BVDV) and identify cattle persistently infected (PI) with this virus, 4,327 samples from Holstein dairy cows were screened over a four-year period in Beijing, China. Eighteen BVD viruses were isolated, 12 from PI cattle. Based on genetic analysis of their 5''-untranslated region (5''-UTR), the 18 isolates were assigned to subgenotype BVDV-1m, 1a, 1d, 1q, and 1b. To investigate the innate immune responses in the peripheral-blood mononuclear cells of PI cattle, the expression of Toll-like receptors (TLRs), RIG-I-like receptors, interferon-α (IFN-α), IFN-β, myxovirus (influenza virus) resistance 1 (MX1), and interferon stimulatory gene 15 (ISG15) was assessed by qPCR. When compared with healthy cattle, the expression of TLR-7, IFN-α, and IFN-β mRNA was downregulated, but the expression of MX1 and ISG-15 mRNA was upregulated in PI cattle. Immunoblotting analysis revealed that the expression of interferon regulatory factor 3 (IRF-3) and IRF-7 was lower in PI cattle than in healthy cattle. Thus, BVDV-1m and 1a are the predominant subgenotypes in the Beijing region, and the strains are highly divergent. Our findings also suggest that the TLR-7/IRF-7 signaling pathway plays a role in evasion of host restriction by BVDV.