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14. Microbiological Profile of the Upper and Lower Respiratory Tract of Suckling and Weaned Dairy Calves with Acute Respiratory Disease.

Author

Alfieri, Alice Fernandes, Fritzen, Juliana Torres Tomazi, Yasumitsu, Carolina Yuka, and Alfieri, Amauri Alcindo

Subjects
BOVINE viral diarrhea virus, AGRICULTURE, ANIMAL herds, HEALTH of cattle, MIXED infections, MYCOPLASMA bovis
Abstract

Simple Summary: Bovine respiratory disease (BRD) is a significant health issue affecting calves in dairy herds worldwide. BRD is caused by viruses and bacteria affecting the upper (URT) and lower respiratory tracts (LRT). The microbiological profile of BRD can vary widely, so it is important to understand the causes of outbreaks to treat and prevent them effectively. This study examined the microbiological profile of nine BRD outbreaks in dairy calves from Paraná, southern Brazil. An analysis was conducted on lung tissue (LRT n = 12) and deep nasopharyngeal swab (URT n = 24) samples. Three herds showed a viral etiology, mainly bovine viral diarrhea virus (BVDV). In three herds, the cause was bacterial (Mycoplasma bovis), while in one herd, a mixed infection of BVDV and M. bovis was found. In two outbreaks, no primary agents for BRD were identified; only bacteria were identified as secondary agents. Viral and single infections were more common in the LRT, causing pneumonia and deaths. Mixed bacterial infections predominated in the URT, involving more than one species of bacteria or bacteria and viruses. In the assessed region, it is recommended that calves receive sufficient levels of passive immunity to BVDV. In cases of BRD, antibiotics targeting M. bovis are recommended for treatment. Bovine respiratory disease (BRD) is a significant global health issue in cattle farming, leading to substantial economic losses. This study analyzed the microbiological profiles of BRD outbreaks in nine dairy cattle herds in southern Brazil. We examined 36 biological samples, including 24 deep nasopharyngeal swabs (NS) and 12 lung tissue, from 29 suckling and 7 weaned heifer calves with acute BRD. PCR and RT-PCR techniques were used to partially amplify the genes of five viruses and four respiratory bacteria. A total of 8 different microorganisms, 4 viruses (bovine viral diarrhea virus, n = 5; bovine coronavirus, n = 3; bovine alphaherpesvirus 1, n = 3; and bovine parainfluenza virus 3, n = 2), and 4 bacteria (Pasteurella multocida, n = 16; Mycoplasma bovis, n = 8; Histophilus somni, n = 7; and Mannheimia haemolytica, n = 4) were identified in 29 (80.5%) samples. Seven samples (four lung tissue and three NS) were negative for all the microorganisms. Mixed infections were more common (62.1%) than single infections (37.9%). Bacterial nucleic acids were more commonly co-detected in NS than in lung tissue. Nucleic acids from a single pathogen were more frequently detected in lung tissues than in NS. M. bovis was the only bacterium detected in the lower respiratory tract. Understanding the microbiological profiles of the respiratory tracts of dairy calves with clinical signs of BRD is crucial for implementing effective biosecurity measures to prevent BRD in suckling and weaned dairy heifer calves. [ABSTRACT FROM AUTHOR]

Published
2024
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15. Higher Frequency of SARS-CoV-2 RNA Shedding by Cats than Dogs in Households with Owners Recently Diagnosed with COVID-19.

Author

Lunardi, Michele, Martins, Felippe Danyel Cardoso, Gustani-Buss, Emanuele, Chideroli, Roberta Torres, de Oliveira, Isabela Medeiros, Peronni, Kamila Chagas, Figueiredo, David Livingstone Alves, Alfieri, Alice Fernandes, and Alfieri, Amauri Alcindo

Subjects
SARS-CoV-2, REVERSE transcriptase polymerase chain reaction, NUCLEOTIDE sequencing, VIRAL genomes, CORONAVIRUSES, PETS
Abstract

Studies have demonstrated the susceptibility of companion animals to natural infection with SARS-CoV-2. Using quantitative reverse transcription polymerase chain reaction and sequencing analyses, this study investigated SARS-CoV-2 RNA excretion in pets in households with infected owners. Oropharyngeal and rectal swabs were collected from dogs and cats in Parana, Southern Brazil, between October 2020 and April 2021. Viral RNA was detected in 25% of cats and 0.98% of dog oropharyngeal swabs; however, systemic, respiratory, and gastrointestinal signs were absent. Complete viral genomes belonged to the Gamma lineage. Phylogenetic analyses indicated that pet samples were probably derived from human-positive cases in Parana. Viral excretion in the oropharynx was more frequent in cats than in dogs. Mutations in the S protein characteristic of Gamma strains were present in all sequenced SARS-CoV-2 strains. The receptor-binding domain of these Brazilian strains did not show any additional mutations not reported in the Gamma strains. Mutations in NSP6, NSP12, and N proteins previously mapped to strains that infect deer or minks were detected. This study highlights the importance of actively monitoring the SARS-CoV-2 strains that infect pets with continued viral exposure. Monitoring genetic changes is crucial because new variants adapted to animals may pose human health risks. [ABSTRACT FROM AUTHOR]

Published
2024
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16. Molecular Detection by Rolling Circle Amplification Combined with Deep Sequencing of Mixed Infection by Bovine Papillomaviruses 2 and 4 in Carcinoma In Situ of the Bovine Esophageal Mucosa.

Author

Matias, Bruna F., Lunardi, Michele, Gonçalves, Kátia C. B., Vilas-Boas, Laurival A., Gustani-Buss, Emanuele, Bracarense, Ana Paula F. R. L., Filho, Luiz Fernando C. Cunha, Alfieri, Alice F., and Alfieri, Amauri A.

Subjects
NUCLEOTIDE sequencing, WHOLE genome sequencing, MIXED infections, ALIMENTARY canal, POLYMERASE chain reaction
Abstract

Papillomaviruses (PVs) are oncogenic and infect the skin and mucosa of various host species. Considering the recent advances in research on PVs using rolling circle amplification (RCA) followed by high-throughput sequencing (HTS), in this study, we aimed to investigate the bovine papillomavirus (BPV) types associated with proliferative lesions in the upper alimentary tract of an affected bull and characterize the viral strains through complete genome sequencing using this strategy. We analyzed the PV strains associated with two hyperplastic esophageal lesions through PCR using degenerate primer pairs and RCA, followed by HTS. HTS of the libraries generated using RCA products provided the whole genome sequence of BPV4 present in squamous papilloma, whereas the complete genome sequence of BPV2 and subgenomic fragments of BPV4 were identified in carcinoma in situ (CIS). For the first time, we have sequenced BPV2 identified from the CIS of the bovine upper alimentary canal. Additionally, RCA followed by HTS allowed characterization of the mixed infection by BPV2 and BPV4 in this lesion. These data reveal that BPV4 is not the only BPV type present in CIS of the esophageal mucous membrane; moreover, a mixed infection caused by BPV2 and BPV4 at the tested anatomical site was demonstrated. [ABSTRACT FROM AUTHOR]

Published
2024
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39. The Role of Mycoplasma bovirhinis in the Development of Singular and Concomitant Respiratory Infections in Dairy Calves from Southern Brazil.

Author

Frucchi, Ana Paula Souza, Dall Agnol, Alais Maria, Caldart, Eloiza Teles, Bronkhorst, Dalton Everton, Alfieri, Alice Fernandes, Alfieri, Amauri Alcindo, and Headley, Selwyn Arlington

Subjects
RESPIRATORY infections, CALVES, ANIMAL herds, MYCOPLASMA, MANNHEIMIA haemolytica, POULTRY farms, DAIRY farm management
Abstract

The role of Mycoplasma bovirhinis in the development of pulmonary disease in cattle is controversial and was never evaluated in cattle from Latin America. This study investigated the respiratory infection dynamics associated with M. bovirhinis in suckling calves from 15 dairy cattle herds in Southern Brazil. Nasal swabs were obtained from asymptomatic (n = 102) and calves with clinical manifestations (n = 103) of bovine respiratory disease (BRD) and used in molecular assays to identify the specific genes of viral and bacterial disease pathogens of BRD. Only M. bovirhinis, bovine coronavirus (BCoV), ovine gammaherpesvirus 2 (OvGHV2), Histophilus somni, Pasteurella multocida, and Mannheimia haemolytica were detected. M. bovirhinis was the most frequently diagnosed pathogen in diseased (57.8%; 59/102) and asymptomatic (55.3%; 57/103) calves at all farms. BCoV-related infections were diagnosed in diseased (52%; 53/102) and asymptomatic (51.4%; 53/103) calves and occurred in 93.3% (14/15) of all farms. Similarly, infectious due to OvGHV2 occurred in diseased (37.2%; 38/102) and asymptomatic (27.2%; /28/103) calves and were diagnosed in 80% (12/15) of all farms investigated. Significant statistical differences were not identified when the two groups of calves were compared at most farms, except for infections due to OvGHV2 that affected five calves at one farm. These results demonstrated that the respiratory infection dynamics of M. bovirhinis identified in Southern Brazil are similar to those observed worldwide, suggesting that there is not enough sufficient collected data to consider M. bovirhinis as a pathogen of respiratory infections in cattle. Additionally, the possible roles of BCoV and OvGHV2 in the development of BRD are discussed. [ABSTRACT FROM AUTHOR]

Published
2024
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43. Molecular Characterization of a New G (VP7) Genotype in Group B Porcine Rotavirus

Author

Molinari, Bruna Letícia Domingues, Alfieri, Alice Fernandes, and Alfieri, Amauri Alcindo

Abstract

Rotaviruses (RVs), a common cause of viral gastroenteritis in humans and animals, are classified into 9 established groups/species (RVA–RVI). Although RVB has been found in several countries, genetic variation among RVB field strains remains poorly characterized. RVB strains can be classified into G genotypes based on a nucleotide (nt) homology that exceeds a cutoff value of 80% for the gene that encodes the structural protein VP7. In this study, we determined the VP7 nt and deduced amino acid sequences of one RVB strain (RB62) identified in a diarrheic fecal sample obtained from a piglet in Brazil in 2012. Comparative analysis of this strain and the strains of the other 21 previously identified VP7 ge­notypes showed that the highest nt identity (71.2%) was found with the porcine PB-70-H5 strain within the G4 genotype. However, when compared with the nonclassified Vietnamese RVB G genotype 14177_18 strain, the nt sequence identity was of 82.9%. These results led us to conclude that the Brazilian strain BR62 and the Vietnamese strain 14177_18 belong to a novel G genotype (G22).

Published
2024
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44. Longitudinal study of rotavirus C VP6 genotype I6 in diarrheic piglets up to 1 week old

Author

Campanha, Joice Elaine Teixeira, Possatti, Flávia, Lorenzetti, Elis, de Almeida Moraes, Daniel, Alfieri, Alice Fernandes, and Alfieri, Amauri Alcindo

Abstract

The reports of rotavirus C (RVC) involvement in diarrhea outbreaks in newborn piglets have been increasing in recent years. This longitudinal study, conducted over a 37-day period, aimed to evaluate the frequency of RVC infection in piglets aged up to 7 days obtained from a pig herd with a previous diagnosis of RVC infection in this age group. Piglets from 50 different litters were monitored daily for the occurrence of diarrhea, and all litters were classified into the following categories: sow parity order (PO) 1 to 5; litter size (LS) ≤ 10 piglets and > 10 piglets; and piglet birth weight (BW) 1.2 to 1.3 kg and > 1.3 to 1.4 kg. Two hundred six diarrheic fecal samples were collected and classified according to the fecal consistency score (pasty, semiliquid, liquid). Ten fecal samples were collected from asymptomatic piglets (control group). Fecal samples were screened for rotavirus (RV) by silver stained-polyacrylamide gel electrophoresis (ss-PAGE), and samples with inconclusive and negative-ss-PAGE results were submitted to RVC VP6 gene amplification by RT-PCR. RVC was identified in 71 (34.5%) samples, in 1 (10%) sample of the control group, and in piglets from 33 (66%) litters. The electrophoretic profile of RV species A was identified in only two samples. Of the 72 RVC-positive samples, 51 (70.8%) presented semiliquid or liquid consistency. There was no significant difference in either group regarding the production parameters (PO, LS, BW) evaluated. An analysis of the whole VP6 gene of three RVC field strains collected on the first, fifteenth, and last day of the experiment enabled us to identify genotype I6. This report describes the first longitudinal study examining epidemiological aspects of RVC infection in newborn piglets.

Published
2024
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