Wild capybaras as reservoir of shiga toxin-producing Escherichia coli in urban Amazonian Region.

Capybaras are rodent widely distributed in South America, which inhabit lakeside areas including ecological parks and urban sites. Due to anthropological interaction, monitoring zoonotic pathogens in wildlife is essential for One Health. We investigated faecal samples from capybaras living in an urban area in Rio Branco (Acre, Brazil) for the presence diarrhoeagenic E. coli. Virulence factors from shiga toxin-producing E. coli (STEC), enterohaemorrhagic E. coli (EHEC), and enteropathogenic E. coli (EPEC) were screened by PCR. We detected at least one virulence factor in 81% of the animals, being classified as STEC and EHEC pathotypes. The presence of zoonotic E. coli in capybaras is a warning due to the highly frequent anthropological interactions with wild animals in this area. Our findings highlight the importance of investigating wild animals as carriers of zoonotic E. coli, requiring further investigations into wildlife surveillance and epidemiological monitoring.

Bovine papillomavirus 24: a novel member of the genus Xipapillomavirus detected in the Amazon region.

Bovine papillomaviruses (BPVs) have been described as etiologic agents of cutaneous and mucosal papillomas in cattle. In the present study, we describe a new BPV that was detected in a cutaneous papilloma from a cow. Phylogenetic analysis suggests that this virus belong to the genus Xipapillomavirus, and we refer to it here as BPV type 24 (BPV24). Coinfection with members of the genera Epsilonpapillomavirus and Deltapapillomavirus in a cutaneous papilloma from a different animal was also detected, and the full genomes of these viruses were sequenced. Both papillomas were from cattle within Acre State in the Amazon region of Brazil. The data presented here demonstrate the utility of using high-throughput methods to indentify coinfections and allow the characterization of new genomes.

Genome characterization of a bovine papillomavirus type 5 from cattle in the Amazon region, Brazil.

Papillomaviruses are small and complex viruses with circular DNA genome that belongs to the Papillomavirus family, which comprises at least 39 genera. The bovine papillomavirus (BPV) causes an infectious disease that is characterized by chronic and proliferative benign tumors that affect cattle worldwide. In the present work, the full genome sequence of BPV type 5, an Epsilonpapillomavirus, is reported. The genome was recovered from papillomatous lesions excised from cattle raised in the Amazon region, Northern Brazil. The genome comprises 7836 base pairs and exhibits the archetypal organization of the Papillomaviridae. This is of significance for the study of BPV biology, since currently available full BPV genome sequences are scarce. The availability of genomic information of BPVs can provide better understanding of the differences in genetics and biology of papillomaviruses.

Complete genome sequence of Deltapapillomavirus 4 (bovine papillomavirus 2) from a bovine papillomavirus lesion in Amazon Region, Brazil.

The complete genome sequence of bovine papillomavirus 2 (BPV2) from Brazilian Amazon Region was determined using multiple-primed rolling circle amplification followed by Illumina sequencing. The genome is 7,947 bp long, with 45.9% GC content. It encodes seven early (E1, E2,E4, E5, E6,E7, and E8) and two late (L1 and L2) genes. The complete genome of a BPV2 can help in future studies since this BPV type is highly reported worldwide although the lack of complete genome sequences available.

Genetic characterization of Amazonian bovine papillomavirus reveals the existence of four new putative types.

Papillomaviruses are small and complex viruses that belong to the Papillomaviridae family, which comprises 39 genera. The bovine papillomavirus (BPV) causes an infectious disease that is characterized by chronic and proliferative benign tumors that affect cattle worldwide. Different genotypes of BPVs can cause distinct skin and mucosal lesions and the immunity they raise has low cross-protection. This report aimed to genotype BPVs in cattle from Northern Brazil based on nucleotide partial sequences of the L1 ORF. Skin wart samples from 39 bovines clinically and histopathologically diagnosed as cutaneous papillomatosis from Acre and Rondônia States were analyzed. The results revealed four already reported BPV types (BPVs 1, 2, 11, and 13), nine putative new BPV subtypes and four putative new BPV types as well as two putative new BPV types that were already reported. To our knowledge, this is the first record of BPVs from the Brazilian Amazon region that identified new possible BPV types and subtypes circulating in this population. These findings point to the great genetic diversity of BPVs that are present in this region and highlight the importance of this knowledge before further studies about vaccination are attempted.

Eyeworms of wild birds and new record of Thelazia (Thelaziella) aquilina (Nematoda: Spirurida).

The eyeworms of the subgenus Thelaziella infect orbital cavities, conjunctival sacs and lachrymal ducts of many wild birds, being able to cause conjunctivitis. In Brazil, at least 10 species of Thelaziella have been described. Here we present a brief review of cases of Thelazia (Thelaziella) aquilina and describe the infection of this parasite in Harpia harpyja, as a new host record. Two nematodes were collected from the surface of the cornea of an adult female H. harpyja in the Amazon region (Brazil) and were morphologically identified as two males of T. (T.) aquilina. This data demonstrated the need for further studies focused on elucidating the transmission, biological cycle, and phylogenetic positioning of Thelaziella as subgenus.

Refrigerated modified Knott concentrate enables long-term morphological viability of canine blood microfilariae.

There are several methods of laboratory diagnosis of filarids, the most used are the thick smear and the Knott method. Both are quick to perform, have a low cost and allow observing the presence, quantifying and analyzing the morphological characteristics of microfilariae. Knowing the morphological viability of fixed microfilariae is of practical importance, as it allows the transport of samples to a laboratory, facilitates epidemiological studies , and allows the storage of samples for didactic. Thus, the aim of this study was to evaluate the morphological viability of microfilariae fixed in the refrigerated modified knott test using 2% formalin. To perform the modified Knott technique, 10 samples of microfilaremic dogs aged over 6 months were used. To evaluate the morphological viability time of the microfilariae in the modified Knott concentrate, the evaluations were repeated after intervals of 0, 1, 7, 30, 60, 120, 180, 240, and 304 days. In the present study, we did not verify any difference in the morphology of the microfilariae in any of the analyzed intervals from day 0 to 304 days, and it is possible to conclude that the use of 2% formalin in the modified Knott technique allows the microfilariae to be identified in a period of 304 days. days after processing the sample without changes in its morphology.

Existem diversos métodos de diagnóstico laboratorial de filarídeos, os mais utilizados são a gota espessa e o método do Knott. Ambos são de rápida execução, possui baixo custo e permitem observar a presença, quantificar e analisar as características morfológicas das microfilárias. Conhecer a viabilidade morfológica das microfilárias fixadas tem importância prática, pois permite o transporte de amostras para um laboratório, facilita estudos epidemiológicos, e permite o armazenamento de amostras para fins didáticos. Desta maneira, o objetivo do trabalho foi avaliar a viabilidade morfológica das microfilárias fixadas em teste do knott modificado refrigerado utilizando formalina a 2%. Para realização da técnica de knott modificado foram utilizadas 10 amostras de cães microfilarêmicos, com idade superior a 6 meses. Para avaliar o tempo de viabilidade morfológica das microfilárias no concentrado de Knott modificado, foram repedidas as avaliações após intervalos de 0,1,7, 30, 60, 120, 180, 240 e 304 dias. No presente estudo não verificamos diferença na morfologia das microfilárias em nenhum dos intervalos analisados deste do dia 0 até 304 dias, sendo possível concluir que a utilização da formalina a 2% na técnica de Knott modificada permite que as microfilárias possam ser identificadas em período de 304 dias após o processamento da amostra sem que ocorram alterações na sua morfologia.

The Development of Novel Primer Sets to Specifically Amplify Each of the Five Different Deltapapillomaviruses That Cause Neoplasia after Cross-Species Infection.

Bovine papillomavirus (BPV) types 1 and 2 are recognized as the main cause of equine sarcoids. However, some studies report that up to a quarter of these tumors do not contain detectible BPV1 or BPV2 DNA. The absence of detectible BPV1 or BPV2 in these sarcoids suggests the possible involvement of other papillomavirus types. Currently, five deltapapillomaviruses are recognized to cause mesenchymal neoplasia after cross-species infection. In addition to BPV1 and BPV2, BPV13 has been associated with equine sarcoids in Brazil, BPV14 has been associated with feline sarcoids, and Ovis aries papillomavirus 2 caused a sarcoid-like lesion in a pig. To investigate the cause of equine sarcoids, PCR primers were developed to specifically amplify each of the five different deltapapillomaviruses that have been associated with mesenchymal neoplasia. The specificity of these primers was confirmed using samples of formalin-fixed tissue known to contain each PV type. These primers allow rapid and sensitive detection of deltapapillomavirus DNA in equine sarcoids. As studies have revealed marked regional variability in the cause of equine sarcoids, these primers will be useful to determine the predominant PV type causing sarcoids in a region. Additionally, there is a single report describing mixed infections by BPV1 and BPV2 in equine sarcoids. The specific primer sets are expected to enable more sensitive detection of mixed infections in equine sarcoids. Determining the cause of equine sarcoids is important as vaccines are developed to prevent these common malignant neoplasms.

The genetic diversity of "papillomavirome" in bovine teat papilloma lesions.

BACKGROUND: Papillomaviruses are small nonenveloped, circular double-stranded DNA viruses that belong to the Papillomaviridae family. To date, 29 Bos taurus papillomavirus (BPV) types have been described. Studies involving mixed BPV infections have rarely been reported in contrast to human papillomavirus (HPV), which is commonly described in numerous studies showing coinfections. Moreover, previous studies had shown that HPV coinfections increase the risk of carcinogenesis. In the present study, we used rolling-circle amplification followed by a high-throughput sequencing (RCA-HTS) approach in 23 teat papillomas from southern Brazil. RESULTS: Eleven well-characterized BPV types and 14 putative new BPV types were genetically characterized into the Xi, Epsilon and Dyoxipapillomavirus genera according to phylogenetic analysis of the L1 gene, which expands the previous 29 BPV types to 43. Moreover, BPV coinfections were detected in the majority (56.3%) of the papilloma lesions analyzed, suggesting a genetic diverse "papillomavirome" in bovine teat warts. CONCLUSIONS: The data generated in this study support the possibility that a wide range of BPV is probably underdetected by conventional molecular detection tools, and that BPV coinfections are underestimated and probably genetic diverse. Additionally, 14 new BPV types were characterized, increasing the knowledge regarding BPV genetic diversity.

Myrciaria dubia Juice (camu-camu) Exhibits Analgesic and Antiedematogenic Activities in Mice.

The Myrciaria dubia (Myrtaceae) fruit is traditionally used to treat malnutrition due to its high levels of vitamin C and phenolic compounds. Because of its composition, this plant is very promising in the research of novel natural treatment for pain disorders. This study analyzed the phytochemical profile of M. dubia juice and assessed its antinociceptive and antiedematogenic potential. The phytochemical profile was determined through high-performance liquid chromatography (HPLC), the oral antinociceptive effect of M. dubia 50% juice (Md50) was evaluated by formalin, hot plate and Complete Freund's Adjuvant tests and the antiedematogenic activity by paw edema. HPLC revealed the presence of ascorbic acid, rutin, and ellagic acid as major compounds. Md50 showed an antinociceptive effect in the acute and chronic phases of the formalin test. In the hot plate test, Md50 also induced an antinociceptive effect of 0.5 up to 6 h, showing antinociceptive and antiedematogenic potential without changing the spontaneous locomotion of animals. All protocols were submitted and approved by the Ethics Committee for use of Animals of the Lutheran University of Brazil (protocol No. 2013-30P).

Trypanosomes of vectors and domestic dogs in Trypanosoma cruzi transmission areas from Brazilian southwestern amazon: New mammalian host for Trypanosoma janseni.

Trypanosoma cruzi is a widespread protozoan in Latin America causing Chagas disease in humans and able to infect several other mammal species. The objective of this study was to investigate the T. cruzi infection in triatomine fauna as well as in dogs from distinct areas of Acre, western Brazilian Amazonia, which recently reported acute cases of human CD as well as an area that have not notify this disease recently. Triatomines were collected and the intestinal contents were evaluated for the presence of trypanosomatids by optical microscopy and polymerase chain reaction (PCR) targeting the mini-exon gene. Blood smear, hemoculture, PCR and serology were performed in the studied mammals. Fecal content of four triatomines were positive (11.6%) in the fresh examination. Molecular analysis identified Trypanosoma cruzi TCI in two specimens. Blood samples from 90 dogs were obtained. Trypanosoma sp. was observed in six blood smears (6/83, 7.22%). Seropositivity for T. cruzi was 8/89 (8.98). One dog's hemoculture was obtained and characterized as T. rangeli. PCR reactions in blood clots resulted in one positive dog (1/75, 1.3%) infected by T. janseni, providing a new mammalian host for a recently described Trypanosoma species. The results demonstrate the low exposition and prevalence for T. cruzi suggesting that dogs are not important to T. cruzi transmission cycle in the studied áreas.

Molecular and pathological characterization of teat papillomatosis in dairy cows in southern Brazil.

Teat papillomatosis is caused by different bovine papillomavirus (BPV) types and is especially important for dairy cows, because it results in severe damage to the health and structure of the mammary gland. This work describes the molecular and pathological aspects of teat papillomatosis in dairy cows in southern Brazil. Samples of teat papillomas were collect from 73 slaughtered dairy cows. Fragments of the lesions were collected in individual pools per animal and subjected to PCR using the FAP primer pair and sequencing of the amplification products. Teats with the remaining lesions were fixed in 10% neutral buffered formalin, routinely processed for histopathology, and stained with hematoxylin and eosin (H&E). Papillomatous lesions were characterized by three macroscopic patterns, namely exophytic (5 [6.9%]), flat (29 [39.7%]), and mixed (39 [53.4%]). Histologically, all samples were identified as squamous papillomas. Partial sequencing of the L1 gene resulted in the detection of 8 classical BPV types (BPVs 4, 6, 7, 8, 9, 10, 11, and 12) in 27 samples, 6 previously reported putative BPV types in 17 samples, and 10 putative new BPV types in 15 samples. Four sequences could not be classified, and 10 were negative in the PCR. There was no correlation between the gross pattern and the BPV type identified, and all the samples were characterized by squamous papillomas under histological examination. However, 24 different BPV types were identified, demonstrating high genetic diversity among BPVs associated with teat papillomatosis in dairy cows in southern Brazil.

Canine papillomavirus type 16 associated to squamous cell carcinoma in a dog: virological and pathological findings.

Papillomaviruses (PVs) are circular double-stranded DNA virus belonging to Papillomaviridae family. During the infection cycle, PVs translate proteins that can influence cell growth and differentiation, leading to epidermal hyperplasia and papillomas (warts) or malignant neoplasms. Canis familiaris papillomaviruses (CPVs) have been associated with different lesions, such as oral and cutaneous papillomatosis, pigmented plaques, and squamous cell carcinomas (SCCs). Here, we report a clinical case of a mixed bred female dog with pigmented plaques induced by CPV16 (Chipapillomavirus 2) that progressed to in situ and invasive SCCs. Gross and histological findings were characterized, and the lesions were mainly observed in ventral abdominal region and medial face of the limbs. In situ hybridization (ISH) revealed strong nuclear hybridization signals in the neoplastic epithelial cells, as well as in the keratinocytes and koilocytes of the pigmented viral plaques. The full genome of the CPV16 recovered directly from the lesions was characterized, and the phylogenetic relationships were determined. The identification of oncoprotein genes (E5, E6, and E7) by high throughput sequencing (HTS) and their expected domains are suggestive of the malignant transformation by CPV16.

How many papillomavirus species can go undetected in papilloma lesions?

A co-infection comprising to at least seven papillomavirus (PV) types was detected by next generation sequencing (NGS) of randomly primed rolling circle amplification (RCA) products of a bovine (Bos taurus) papilloma lesion from the Brazilian Amazon region. Six putative new PV types that could not be detected by commonly used PCR protocols were identified. Their overall L1 nucleotide identities were less than 90% compared to described PV species and types. L1 nucleotide BLAST sequence hits showed that each new type was related to Beta, Gamma, Dyokappa, Dyoeta, and Xipapillomavirus, as well as two likely new unclassified genera. Our results show that the employment of NGS is relevant to the detection and characterization of distantly related PV and is of major importance in co-infection studies. This knowledge will help us understand the biology and pathogenesis of PV, as well as contribute to disease control. Moreover, we can also conclude that there are many unknown circulating PVs.

Novel Bovine Papillomavirus Type Discovered by Rolling-Circle Amplification Coupled with Next-Generation Sequencing.

Currently, fifteen bovine papillomavirus (BPV) types have been identified and classified into four genera: Deltapapillomavirus, Epsilonpapillomavirus, Dyoxipapillomavirus, and Xipapillomavirus. Here, the complete genome sequence of a new BPV type (BPV 04AC14) recovered from a papillomatous lesion is reported. The genome is 7,282 bp in length and exhibits the classic genetic organization and motifs of the members of Papillomaviridae. Maximum likelihood phylogenetic analyses revealed that BPV 04AC14 clusters with members of the Xipapillomavirus genus. The nucleotide sequence of the L1 capsid protein of the novel BPV is closely related to its counterpart, BPV3, with which it shares 79% similarity. These findings suggest that this virus is a new BPV type of the Xipapillomavirus genus.