Report of Giardia duodenalis in a non-captive chital Axis axis (Erxleben 1777) in Santa Catarina, South of Brazil.

Giardiasis is an infection of the small intestine by protozoa of the genus Giardia, which has a wide range of susceptible hosts, including domestic and wild animals and humans. Giardia is a zoonotic agent and represents one of the main human parasites, with high prevalence and great importance in public health. This report aims to describe the parasitism of a non-captive Chital deer (Axis axis) by Giardia duodenalis. The animal, after being rescued by soldiers from the 2nd Platoon of the 2nd Company of the 2nd Battalion of the Environmental Military Police of the State of Santa Catarina, was sent to the Wildlife Studies Center (NEVS) of the University of Western Santa Catarina (UNOESC). During clinical care, an exposed fracture in the left pelvic limb and signs of acute respiratory failure were found. Fecal samples were collected for later parasitological diagnosis. Two techniques were applied: centrifugal flotation with zinc sulfate, to diagnose parasites of the gastrointestinal system, and Baermann, to search for parasitism in the respiratory tract. The investigation revealed the presence of Giardia duodenalis. The animal died on the same day of its arrival due to a cardiorespiratory arrest. The presence of this parasite in an invasive exotic deer species highlights its epidemiological importance, as it can act as a source of infection and spread the disease to humans and other animals.

Giardíase é uma infecção do intestino delgado por protozoários do gênero Giardia, o qual apresenta uma ampla variedade de hospedeiros suscetíveis, incluindo animais domésticos, selvagens e também humanos. Giardia é um agente zoonótico, e representa um dos principais parasitas de humanos, com alta prevalência e grande importância em saúde pública. Este relato tem como objetivo descrever o parasitismo de um indivíduo de cervo Chital (Axis axis) de vida livre por Giardia duodenalis. O animal, após ser resgatado por soldados do 2º Pelotão da 2ª Companhia do 2º Batalhão da Polícia Militar Ambiental do Estado de Santa Catarina, foi encaminhado ao Núcleo de Estudos em Vida Selvagem (NEVS) da Universidade do Oeste de Santa Catarina (UNOESC). Durante o atendimento clínico foi constatada fratura exposta no membro pélvico esquerdo e sinais de insuficiência respiratória aguda. Amostras de fezes foram coletadas para posterior diagnóstico parasitológico. Para isso, foram usadas duas técnicas: centrífugo-flutuação em sulfato de zinco, com o objetivo de diagnosticar parasitas do sistema gastrointestinal, e Baermann, com o objetivo de pesquisar parasitas do trato respiratório. Os achados revelaram presença de cistos de Giardia duodenalis. O animal morreu no mesmo dia de sua chegada, devido a uma parada cardiorrespiratória. A presença desse parasito em uma espécie de cervídeo exótico invasor destaca sua importância epidemiológica, pois o mesmo pode atuar como fonte de infecção e de disseminação da doença para o ser humano e/ou outros animais.

PRESENCE OF ANTI-NEOSPORA CANINUM ANTIBODIES IN TEMAZATE DEER (MAZAMA TEMAMA) AND WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) FROM THE CENTRAL REGION OF VERACRUZ, MEXICO.

This study documents the presence of anti-Neospora caninum antibodies and their association with certain risk factors in 2 deer species from the central region of Veracruz State, Mexico. A total of 90 blood samples, 20 from temazate deer (Mazama temama) and 70 from white-tailed deer (Odocoileus virginianus), were taken from 3 farms, and serum samples were subjected to ELISA indirect test to detect N. caninum antibodies; the association between the serological status and the possible risk factors was then estimated. The overall presence of anti-N. caninum antibodies was 57.7% (52/90; 95% CI 46.9-67.9), with positive animals identified on all farms; in white-tailed deer it was 57% and in temazate deer 60%. Prevalence was higher in females than males. Adult animals had a higher prevalence than young ones. The risk analysis identified the age in the adult animal category (odds ratio 5.8) as being associated with the presence of anti-N. caninum antibodies. These results provide evidence of the significant contamination of oocysts in the environment and allow us to estimate the contribution of deer to the sylvatic cycle.

Comparative karyotype analysis of the red brocket deer (M. americana sensu lato and M. rufa) complex: evidence of drastic chromosomal evolution and implications on speciation process.

Chromosomal rearrangements are often associated with playing a role in the speciation process. However, the underlying mechanism that favors the genetic isolation associated with chromosomal changes remains elusive. In this sense, the genus Mazama is recognized by its high level of karyotype diversity among species with similar morphology. A cryptic species complex has been identified within the genus, with the red brocket deer (Mazama americana and Mazama rufa) being the most impressive example. The chromosome variation was clustered in cytotypes with diploid numbers ranging from 42 to 53 and was correlated with geographical location. We conducted an analysis of chromosome evolution of the red brocket deer complex using comparative chromosome painting and Bacterial Artificial Chromosome (BAC) clones among different cytotypes. The aim was to deepen our understanding of the karyotypic relationships within the red brocket, thereby elucidating the significant chromosome variation among closely related species. This underscores the significance of chromosome changes as a key evolutionary process shaping their genomes. The results revealed the presence of three distinct cytogenetic lineages characterized by significant karyotypic divergence, suggesting the existence of efficient post-zygotic barriers. Tandem fusions constitute the main mechanism driving karyotype evolution, following a few centric fusions, inversion X-autosomal fusions. The BAC mapping has improved our comprehension of the karyotypic relationships within the red brocket deer complex, prompting questions regarding the role of these changes in the speciation process. We propose the red brocket as a model group to investigate how chromosomal changes contribute to isolation and explore the implications of these changes in taxonomy and conservation.

Diversity of midgut microbiota in ticks collected from white-tailed deer (Odocoileus virginianus) from northern Mexico.

Ticks host different pathogens as endosymbiont and nonpathogenic microorganisms and play an important role in reproductive fitness and nutrient provision. However, the bacterial microbiomes of white-tailed deer ticks have received minimal attention. This study aimed to examine the bacterial microbiome of ticks collected from Odocoileus virginianus on the Mexico-United States border to assess differences in microbiome diversity in ticks of different species, sexes, and localities. Five different tick species were collected: Rhipicephalus microplus, Dermacentor nitens, Otobius megnini, Amblyomma cajennense, and A. maculatum. The tick microbiomes were analyzed using next-generation sequencing. Among all tick species, the most predominant phylum was Proteobacteria, followed by Actinobacteria and Firmicutes. The ticks from Tamaulipas and Nuevo León presented the highest bacterial species diversity. Acinetobacter johnsonii and A. lwoffii were the common bacterial species in the microbiome of all ticks, Coxiella were present in R. microplus, and Dermacentor nitens also exhibited a Francisella-like endosymbiont. The microbiome of most females in D. nitens was less diverse than that of males, whereas R. microplus occurs in females, suggesting that microbiome diversity is influenced by sex. In the bacterial communities of A. maculatum and O. megnini, Candidatus Midichloria massiliensis, and Candidatus Endoecteinascidia fumentensis were the most predominant endosymbionts. These results constitute the initial report on these bacteria, and this is also the first study to characterize the microbiome of O. megnini.

The Study of Bluetongue Virus (BTV) and Epizootic Hemorrhagic Disease Virus (EHDV) Circulation and Vectors at the Municipal Parks and Zoobotanical Foundation of Belo Horizonte, Minas Gerais, Brazil (FPMZB-BH).

Bluetongue Virus (BTV) and Epizootic Hemorrhagic Disease Virus (EHDV) are Orbiviruses primarily transmitted by their biological vector, Culicoides spp. Latreille, 1809 (Diptera: Ceratopogonidae). These viruses can infect a diverse range of vertebrate hosts, leading to disease outbreaks in domestic and wild ruminants worldwide. This study, conducted at the Belo Horizonte Municipal Parks and Zoobotany Foundation (FPMZB-BH), Minas Gerais, Brazil, focused on Orbivirus and its vectors. Collections of Culicoides spp. were carried out at the FPMZB-BH from 9 December 2021 to 18 November 2022. A higher prevalence of these insects was observed during the summer months, especially in February. Factors such as elevated temperatures, high humidity, fecal accumulation, and proximity to large animals, like camels and elephants, were associated with increased Culicoides capture. Among the identified Culicoides spp. species, Culicoides insignis Lutz, 1913, constituted 75%, and Culicoides pusillus Lutz, 1913, 6% of the collected midges, both described as competent vectors for Orbivirus transmission. Additionally, a previously unreported species in Minas Gerais, Culicoides debilipalpis Lutz, 1913, was identified, also suspected of being a transmitter of these Orbiviruses. The feeding preferences of some Culicoides species were analyzed, revealing that C. insignis feeds on deer, Red deer (Cervus elaphus) and European fallow deer (Dama dama). Different Culicoides spp. were also identified feeding on humans, raising concerns about the potential transmission of arboviruses at the site. In parallel, 72 serum samples from 14 susceptible species, including various Cervids, collected between 2012 and 2022 from the FPMZB-BH serum bank, underwent Agar Gel Immunodiffusion (AGID) testing for BTV and EHDV. The results showed 75% seropositivity for BTV and 19% for EHDV. Post-testing analysis revealed variations in antibody presence against BTV in a tapir and a fallow deer and against EHDV in a gemsbok across different years. These studies confirm the presence of BTV and EHDV vectors, along with potential virus circulation in the zoo. Consequently, implementing control measures is essential to prevent susceptible species from becoming infected and developing clinical diseases.

Phylogenetic congruence of Plasmodium spp. and wild ungulate hosts in the Peruvian Amazon.

Malaria parasites are known to infect a variety of vertebrate hosts, including ungulates. However, ungulates of Amazonia have not been investigated. We report for the first time, the presence of parasite lineages closely related to Plasmodium odocoilei clade 1 and clade 2 in free-ranging South American red-brocket deer (Mazama americana; 44.4%, 4/9) and gray-brocket deer (Mazama nemorivaga; 50.0%, 1/2). We performed PCR-based analysis of blood samples from 47 ungulates of five different species collected during subsistence hunting by an indigenous community in the Peruvian Amazon. We detected Plasmodium malariae/brasilianum lineage in a sample from red-brocket deer. However, no parasite DNA was detected in collared peccary (Pecari tajacu; 0.0%, 0/10), white-lipped peccary (Tayassu pecari; 0.0%, 0/15), and tapir (Tapirus terrestris; 0.0%, 0/11). Concordant phylogenetic analyses suggested a possible co-evolutionary relationship between the Plasmodium lineages found in American deer and their hosts.

Anatomy and computed tomography of the nasal cavity, nasal conchae, and paranasal sinuses of the endangered Patagonian huemul deer (Hippocamelus bisulcus).

This study explores for the first time the shape, volume, and configuration of nasal cavity structures of the endangered Patagonian huemul deer via computed tomography (CT). Three-dimensional (3D) reconstructions derived from data sets obtained from five Patagonian huemul deer skulls were analyzed. Using semiautomatic segmentation, 3D models were created of all the sinus compartments and nasal conchae. Volumetric measurements were taken of seven sinus compartments. The Patagonian huemul deer has a wide, large nasal cavity, with an osseous nasal aperture typical of cervids and a choana with characteristics that differentiate it from the pudu and roe deer. It also has six nasal meatuses and three nasal conchae, with the ventral nasal concha having the greatest volume and surface, which given its extension ensures a greater ability to humidify and heat the air. Further analysis showed the complex system of paranasal sinuses to be characterized by a rostroventral and interconnected group, where communication with the nasal cavity is common through the nasomaxillary opening, and a caudodorsal group that communicates with the nasal cavity through openings in the nasal meatuses. Our study of the endangered Patagonian huemul deer documents an intricate, and in some nasal cavity structures, unique morphological construction which may predispose it to higher rates of sinonasal afflictions due largely to its nasal complex anatomy, thus affecting its high cultural value.

A Method to Freeze Skin Samples for Cryobanks: A Test of Some Cryoprotectants for an Endangered Deer.

The genetic diversity of endangered deer species, such as Mazama jucunda, can be preserved with the help of somatic cell cryopreservation. This procedure allows obtaining several cells from the individual even after its death, which is very important for applications in reproductive biotechnologies. This study's objective was to test cryopreservation protocols of skin fragments of M. jucunda, using different cryoprotectants in slow freezing. We evaluated four treatments, composed of three cryoprotectants, dimethyl sulfoxide (DMSO), polyvinylpyrrolidone (PVP), and ethylene glycol (EG), used alone and in combination. There was also a control group where the tissue did not undergo cryopreservation. Skin fragments were collected from the medial region of the pelvic limbs of three individuals. Each fragment was divided into 10 equal parts, standardized by weight, making two pieces for each treatment and control from each animal. The collected fragments were evaluated in culture, based on the speed of occupation of the free spaces of the cell culture flask. Cell viability was also evaluated using Trypan Blue dye and the mitotic index to understand the effect of toxicity and freezing on cell membrane integrity and cell division capacity, respectively. The treatments that used association with PVP proved to be more damaging to the cells, taking longer to reach confluence. EG alone showed better results than DMSO in the slow-freezing protocol. Clinical Trial Registration Number is 1390/21.

Cervids ACE2 Residues that Bind the Spike Protein can Provide Susceptibility to SARS-CoV-2.

The susceptibility of the white-tailed deer (WTD; Odocoileus virginianus) to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted cervids as coronavirus reservoirs. This study aimed to evaluate the angiotensin-converting enzyme 2 (ACE2) residues which bind the spike protein of SARS-CoV-2 from 16 cervids to predict their potential susceptibility to SARS-CoV-2 infection. Eleven out of 16 species presented identical ACE2 key residues to WTD ACE2. Four cervids presented K31N, a variant associated with low SARS-CoV-2 susceptibility. Large herding of cervids with ACE2 key residues identical to that of the WTD can result in extensive reservoirs of SARS-CoV-2. Cervids as potential reservoirs could favor SARS-CoV-2 adaptation and the emergence of new coronavirus strains.

Identification of a new Sarcocystis sp. in marsh deer (Blastocerus dichotomus) from wetlands of Argentina.

The marsh deer (Blastocerus dichotomus) is the largest South American native deer species and is listed as "Vulnerable" by IUCN due to the population reduction. As part of a conservation and disease surveillance program, muscle samples from 14 marsh deer found dead in 2016 and 2017 in northeast Argentina were obtained at necropsy. Samples from each animal were processed as pooled muscles (heart, diaphragm, tongue and hindlimb) by homogenization and direct microscopical observation to detect intracellular Sarcocystis spp. cysts. Sarcocysts were observed in six samples, and several cysts recovered from two samples were processed by transmission electron microscopy. The cysts were thin-walled and showed a cyst-wall ultrastructure with ribbon-like protrusions similar to other species using cervids as intermediate host and canids as definitive hosts. Genomic DNA from individual sarcocysts from three marsh deer were successfully amplified by PCR of 18S rRNA and COI gene fragments and further sequenced. Sequence comparison revealed a 99.3-100% identity among them and only 93.7-96.6% and 88.8-89.7% identity at 18S rRNA and COI markers, respectively, with other Sarcocystis spp. Despite morphological similarities, the high sequence divergence at 18S rRNA and COI fragments allowed the assumption that Sarcocystis sp. from marsh deer is a different species from others using cervids as intermediate hosts. Therefore, we propose the name Sarcocystis blastoceris n. sp. for the species infecting marsh deer.

Quasi-static and dynamic Brazilian testing and failure analysis of a deer antler in the transverse to the osteon growth direction.

The transverse tensile strength of a naturally fallen red deer antler (Cervus Elaphus) was determined through indirect Brazilian tests using dry disc-shape specimens at quasi-static and high strain rates. Dynamic Brazilian tests were performed in a compression Split-Hopkinson Pressure Bar. Quasi-static tensile and indirect Brazilian tests were also performed along the osteon growth direction for comparison. The quasi-static transverse tensile strength ranged 31.5-44.5 MPa. The strength increased to 83 MPa on the average in the dynamic Brazilian tests, proving a rate sensitive transverse strength. The quasi-static tensile strength in the osteon growth direction was however found comparably higher, 192 MPa. A Weibull analysis indicated a higher tensile ductility in the osteon growth direction than in the transverse to the osteon growth direction. The microscopic analysis of the quasi-static Brazilian test specimens (tensile strain along the osteon growth direction) revealed a micro-cracking mechanism operating by the crack deflection/twisting at the lacunae in the concentric lamellae region and at the interface between concentric lamellae and interstitial lamellae. On the other side, the specimens in the transverse direction fractured in a more brittle manner by the separation/delamination of the concentric lamellae and pulling of the interstitial lamellae. The detected increase in the transverse strength in the high strain rate tests was further ascribed to the pull and fracture of the visco-plastic collagen fibers in the interstitial lamellae. This was also confirmed microscopically; the dynamically tested specimens exhibited flatter fracture surfaces.

Functional characterization of compound heterozygosity Hb S/Hb Deer Lodge in Brazil.

INTRODUCTION: The Hb Deer Lodge (ß2 His>Arg; HBB:c.8A>G) is a structural hemoglobin variant described in some populations around the world, characterized by increased oxygen affinity, but does not confer clinical symptoms to its carriers. The coinheritance of the Hb Deer Lodge with the most common hemoglobin variant, Hb S, has been reported only once; however, functional data were not described. Here we show a case of the Hb S and Hb Deer Lodge carrier in heterozygosity. METHODS: The Hb S and Hb Deer Lodge association was identified by High-Performance Liquid Chromatography (HPLC), reverse phase HPLC and the ß globin gene sequencing. The functional characterization of this interaction was obtained using the O2 dissociation curve, determination of the cooperativity between the globin chains and the Bohr effect in the presence and absence of organic phosphates. RESULTS: When the Hb S and Hb Deer Lodge were associated, there was a decrease in cooperativity, no significant changes in oxygen affinity and no significant Bohr effect changes. CONCLUSION: Despite these genetic variations, the carrier showed no hematological alterations and no clinical symptoms, possibly due to the high oxygen affinity of the Hb Deer Lodge, which interferes with the Hb S polymerization.

Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp. ) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome / Semelhanças genéticas e análise filogenética de muntjac (Muntiacus spp. ) comparando a sequência de nucleótidos de rRNA 16S e genoma citocromo B

This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.(AU)

Este estudo visou identificar as semelhanças filogenéticas entre os muntjac (Muntiacus spp.). As semelhanças filogenéticas entre sete grandes espécies muntjac foram estudadas comparando a sequência de nucleótidos de 16s rRNA e genoma citocromo b. As sequências de nucleótidos, obtidas a partir de bases de dados NCBI, foram alinhadas utilizando o software DNASTAR. Foi criada uma árvore filogenética para as espécies selecionadas de muntjac utilizando o método de probabilidade máxima no software MEGA7. Os resultados das sequências de nucleótidos (16s rRNA) mostraram semelhanças filogenéticas entre o M. truongsonensis e o M. rooseveltorum tiveram o maior número (99,2%) enquanto as semelhanças mais baixas (96,8%) encontradas entre M. crinifrons e M. putaoensi. Enquanto os resultados das sequências de nucleótidos (Cty-b) apresentaram a maior semelhança (100%) entre M. muntjak e M. truongsonensis e os mais baixos (91,5%) entre M. putaoensis e M. crinifrons. A árvore filogenética das espécies muntjac (gene rRNA 16s) mostra os dois principais aglomerados, o que inclui M. putaoensis, M. truongsonensis, M. rooseveltorum e M. muntjak, e o segundo incluindo M. crinifrons e M. vuquangensis. O M. reevesi existe separadamente na árvore filogenética. A árvore filogenética das espécies muntjac usando genes citocromo b mostra que os M. muntjak e M. truongsonensis estão agrupados no mesmo grupo.(AU)

Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp. ) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome / Semelhanças genéticas e análise filogenética de muntjac (Muntiacus spp. ) comparando a sequência de nucleótidos de rRNA 16S e genoma citocromo B

This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.

Este estudo visou identificar as semelhanças filogenéticas entre os muntjac (Muntiacus spp.). As semelhanças filogenéticas entre sete grandes espécies muntjac foram estudadas comparando a sequência de nucleótidos de 16s rRNA e genoma citocromo b. As sequências de nucleótidos, obtidas a partir de bases de dados NCBI, foram alinhadas utilizando o software DNASTAR. Foi criada uma árvore filogenética para as espécies selecionadas de muntjac utilizando o método de probabilidade máxima no software MEGA7. Os resultados das sequências de nucleótidos (16s rRNA) mostraram semelhanças filogenéticas entre o M. truongsonensis e o M. rooseveltorum tiveram o maior número (99,2%) enquanto as semelhanças mais baixas (96,8%) encontradas entre M. crinifrons e M. putaoensi. Enquanto os resultados das sequências de nucleótidos (Cty-b) apresentaram a maior semelhança (100%) entre M. muntjak e M. truongsonensis e os mais baixos (91,5%) entre M. putaoensis e M. crinifrons. A árvore filogenética das espécies muntjac (gene rRNA 16s) mostra os dois principais aglomerados, o que inclui M. putaoensis, M. truongsonensis, M. rooseveltorum e M. muntjak, e o segundo incluindo M. crinifrons e M. vuquangensis. O M. reevesi existe separadamente na árvore filogenética. A árvore filogenética das espécies muntjac usando genes citocromo b mostra que os M. muntjak e M. truongsonensis estão agrupados no mesmo grupo.

Genetic similarities and phylogenetic analysis of Muntjac (Muntiacus spp.) by comparing the nucleotide sequence of 16S rRNA and cytochrome B genome

Abstract This study aimed to identify the phylogenetic similarities among the muntjac (Muntiacus spp.). The phylogenetic similarities among seven major muntjac species were studied by comparing the nucleotide sequence of 16s rRNA and cytochrome b genome. Nucleotide sequences, retrieved from NCBI databases were aligned by using DNASTAR software. A phylogenetic tree was created for the selected species of muntjac by using the maximum likelihood method on MEGA7 software. The results of nucleotide sequences (16s rRNA) showed phylogenetic similarities between, the M. truongsonensis and M. rooseveltorum had the highest (99.2%) while the lowest similarities (96.8%) found between M. crinifrons and M. putaoensi. While the results of nucleotide sequences (Cty b) showed the highest similarity (100%) between M. muntjak and M. truongsonensis and the lowest s (91.5%) among M. putaoensis and M. crinifrons. The phylogenetic tree of muntjac species (16s rRNA gene) shows the main two clusters, the one including M. putaoensis, M. truongsonensis, M. rooseveltorum, and M. muntjak, and the second one including M. crinifrons and M. vuquangensis. The M. reevesi exists separately in the phylogenetic tree. The phylogenetic tree of muntjac species using cytochrome b genes shows that the M. muntjak and M. truongsonensis are clustered in the same group.

Resumo Este estudo visou identificar as semelhanças filogenéticas entre os muntjac (Muntiacus spp.). As semelhanças filogenéticas entre sete grandes espécies muntjac foram estudadas comparando a sequência de nucleótidos de 16s rRNA e genoma citocromo b. As sequências de nucleótidos, obtidas a partir de bases de dados NCBI, foram alinhadas utilizando o software DNASTAR. Foi criada uma árvore filogenética para as espécies selecionadas de muntjac utilizando o método de probabilidade máxima no software MEGA7. Os resultados das sequências de nucleótidos (16s rRNA) mostraram semelhanças filogenéticas entre o M. truongsonensis e o M. rooseveltorum tiveram o maior número (99,2%) enquanto as semelhanças mais baixas (96,8%) encontradas entre M. crinifrons e M. putaoensi. Enquanto os resultados das sequências de nucleótidos (Cty-b) apresentaram a maior semelhança (100%) entre M. muntjak e M. truongsonensis e os mais baixos (91,5%) entre M. putaoensis e M. crinifrons. A árvore filogenética das espécies muntjac (gene rRNA 16s) mostra os dois principais aglomerados, o que inclui M. putaoensis, M. truongsonensis, M. rooseveltorum e M. muntjak, e o segundo incluindo M. crinifrons e M. vuquangensis. O M. reevesi existe separadamente na árvore filogenética. A árvore filogenética das espécies muntjac usando genes citocromo b mostra que os M. muntjak e M. truongsonensis estão agrupados no mesmo grupo.

Skull anatomy of the endangered Patagonian huemul deer(Hippocamelus bisulcus).

Studies of the normal skull anatomy of the Patagonian Huemul deer are scarce. Currently, the findings of bone lesions in the skull associated with metabolic imbalances are frequent in the literature. The objective of this study was to provide anatomical and morphometric data of the cranium and facies including a morphofunctional interpretation as a reference for clinical, ecomorphological and educational purposes. Five skulls were described, measured, scanned and digitally reconstructed. The presence of a caudal projection of the vomer bone, the absence of the facial tubercle and thin bones forming the cranial cavity were observed. Linear measurements allowed the skull to be classified as dolichocephalic, hyperlepten and ultra-dolichocranial. In conclusion, the Patagonian Huemul has a long head, an extremely long skull and a very narrow face. The thickness of the bones that made up the walls of the cranial cavity suggests chronic metabolic imbalances in response to mineral deficiency. The anatomical and morphometric data obtained in this study strongly emphasizes the relevance of considering the implementation of such management policies that aim at promoting an optimal nutritional context.

Epidemic abortions due to Neospora caninum infection in farmed red deer (Cervus elaphus).

This study describes for the first time an abortion outbreak caused by Neospora caninum in farmed red deer. During a 5-year period, farmed hinds, naturally mated, were regularly ultrasound monitored to detect reproductive losses over their gestation. During the 4 years previous to the outbreak, abortion rates ranged from 4.7 to 8.6% (average 6.5%), and serology for indirect diagnosis of neosporosis and toxoplasmosis was performed. At the fifth year, the abortion rate increased to 25.3%. During this outbreak, three aborted foetuses and their placentas were recovered and submitted to laboratory for etiological diagnosis. Blood samples were collected from the 81 hinds at the end of the gestational period and the seropositivity rate for N. caninum, Toxoplasma gondii, Brucella abortus, bovine viral diarrhoea virus and bovine alphaherpesvirus type 1 was 66.7%, 67.9%, 0.0%, 8.6% and 0.0%, respectively. Neospora caninum-seropositive hinds (OR = 5.7, P = 0.0271) and hinds with high antibody titres to N. caninum (OR = 7.4, P = 0.0130) were more likely to abort than seronegative hinds. In addition, N. caninum seropositivity rate in the aborted hinds was higher (OR = 5.4, P = 0.033) than the non-aborted hinds. No association was found between T. gondii nor BVDV-seropositivity and abortions. Typical protozoal histopathologic findings (necrotizing non suppurative encephalitis, meningitis, myocarditis, hepatitis, among others) were observed in all foetuses. Neospora caninum was immunolabelled by immunohistochemistry in several tissues from two foetuses, and infection was also confirmed in the three foetuses by serology and/or DNA detection. No other abortifacient agent was detected in the foetuses. Their dams showed high N. caninum antibody titres (≥ 6400). Serologic evidence and epidemiological data recorded suggested a point-source of N. caninum infection before the occurrence of the outbreak, probably related with contaminated feedstuff with oocysts. Moreover, the intensive production system with a high stocking rate could be also considered a factor which might have increased the risk of horizontal N. caninum infection in this herd.

Characteristics of Central American brocket deer resting sites in a tropical mountain cloud forest in eastern Mexico.

The Central American brocket deer is a vulnerable species. Geographically isolated populations have been affected by poaching and habitat fragmentation, leading to local extinctions. It is therefore important to understand this species' habitat characteristics, particularly of resting sites, which play a crucial role in survival and fitness. We describe the characteristics and distribution patterns of Central American brocket deer resting sites at the microhabitat and landscape scales in San Bartolo Tutotepec, Hidalgo, México. We conducted eight bimonthly field surveys between November 2017 and March 2019, consisting of 32 transects of 500 m length to search for fecal pellets, footprints, scrapes, and browsed plants. At each resting site we identified, we measured canopy closure, horizontal thermal cover, protection from predators for fawns and adults, escape routes, slope from the ground, presence of scrapes, cumulative importance value of the edible plant species, and distance from the resting site to the nearest water resource to characterize the site at the microhabitat scale. At the landscape scale, we identified the type of biotope, elevation, aspect, and slope. We compared all of these parameters from resting sites with a paired randomly selected site to serve as a control. We performed a multiple logistic regression to identify the parameters associated with the resting sites and a point pattern analysis to describe their distribution. We characterized 43 resting sites and their corresponding control plots. At the microhabitat scale, resting sites were associated with higher vertical thermal cover, more concealment cover, more escape routes, more edible plant species, higher slope from the ground, and closer distance to water resources. At the landscape scale, resting sites were associated with beech forest, oak forest, secondary forest, and ravine biotopes and negatively associated with pine forest, houses, and roads. Resting sites had an aggregated spatial pattern from 0 to 900 m, but their distribution was completely random at larger scales. Our study revealed that Central American brocket deer selected places with specific characteristics to rest, at both microhabitat and landscape scales. We therefore suggest that existing habitat be increased by reforesting with native species-particularly Mexican beech forest and oak forest-to improve the deer's conservation status in the study area.

First report of Lipoptena mazamae Rondani, 1878 (Diptera: Hippoboscidae) from Paraguay.

We report for the first time from Paraguay the louse-fly Lipoptena mazamae Rondani, 1878 (Diptera: Hippoboscidae), based on an individual retrieved from the body of a gray brocket Mazama gouazoubira (Fischer), found dead in the Tatí Yupí Biological Refuge, in Alto Paraná, Eastern Paraguay. Before this finding, in the decade of the 1940's, this species of fly was wrongly recorded from Paraguay, based on material collected in Bolivia, some 300 km from the border to Paraguay.

FIRST REPORT OF GENUS Cryptosporidium IN CERVIDS SPECIES: Mazama americana, Mazama nana AND Blastocerus dichotomus.

We analyzed Cryptosporidium spp. in fecal samples of wild cervids (Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo) from many Brazilian regions, a fact unprecedented in the literature. Sniffer dogs were used to collect 936 fecal samples of cervids from 14 Brazilian localities. Cervids species were identified using polymerase chain reaction (PCR) performed from genomic DNA extracted from 563 fecal samples of Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo. Cryptosporidium spp. oocyst screening was performed using malachite green negative staining. Nested PCR (nPCR) protocols targeting the 18S rRNA and GP60 genes followed by genetic sequencing were performed for Cryptosporidium spp. detection and Cryptosporidium parvum subtyping, respectively. Nested PCR targeting actin gene and genetic sequencing were performed in samples with non-identified Cryptosporidium species by 18S rRNA amplicon sequencing. The association between the occurrence of Cryptosporidium and the presence of bovines in the same locality was evaluated using Fisher's exact test. The positivity rates of diagnostic methods were compared by McNemar test and the Kappa correlation coefficient. The prevalence rates of Cryptosporidium spp. in cervids were 1.42% (8/563) and 0.36% (2/563) by nPCR and malachite green negative staining, respectively. C. parvum IIaA16G3R1 isolate was identified in three fecal samples from M. americana, two from M. nana and one from B. dichotomus. Cryptosporidium ryanae were found in one sample from B. dichotomus. We identified a new Cryptosporidium genotype, named Cryptosporidium deer genotype BR, from one M. americana fecal sample.