Investigating the need for antibiotic supplementation to the extender used for semen cryopreservation in collared peccaries.

The objective was to investigate the effects of semen freezing extender supplementation with antibiotics on bacterial load of semen samples, sperm functional and morphological metrics in the collared peccary. Fresh ejaculates from 10 males were extended in Tris-egg yolk-glycerol supplemented or not (control) with gentamicin (70 µg/mL) streptomycin-penicillin (SP; 1 mg/mL-1000 IU/mL) or and cryopreserved in liquid nitrogen. Bacterial load, sperm motility patterns, morphology, membrane functionality and integrity, mitochondrial activity, chromatin integrity and sperm-binding ability were evaluated in fresh and frozen-thawed samples. Regardless of the use of antibiotics, the sole cryopreservation provoked a significant decrease (P < 0.05) in bacterial load compared to fresh samples (from average values > 1 x 106 CFU/mL to <0.4 × 106 CFU/mL). Post-thawing sperm kinetic parameters were not affected by the absence or presence of different antibiotics, except for beat cross frequency that was significantly (P < 0.05) impaired by SP supplementation compared to the group without antibiotics. After thawing, sperm morphology, membrane functionality and integrity, and mitochondrial activity were also not affected by the presence or absence of antibiotics; however, a significant decrease was observed in the group without antibiotics (P < 0.05) in comparison to fresh samples. Regarding sperm-binding ability, there were no differences among the different groups. While collared peccary semen could be efficiently cryopreserved in the absence of antibiotics in the extender, the use of both gentamicin or the streptomycin-penicillin combination is recommended as effective antibiotic supplementation for a further control of bacterial loads without affecting sperm parameters.

Embryonic and fetal development of the collared peccary (Pecari tajacu).

The relative timing of the main fetal development events in species determine the extent of fetal development at birth, which range along a gradient of having altricial and precocial traits. The results from this study allow for description of important fetal developments in collared peccary (Pecari tajacu) using data from 118 embryo/fetuses from 68 pregnant peccaries obtained over a period of 15 years through collaborative methods with local hunters in the Amazon. The chronological order of emergence of external characteristics in relation to the total dorsal length (TDL) was: differentiated genitalia, limbs and eyelid buds (TDL ≥ 4.5 cm), fusioned eyelids and outer ear (TDL ≥ 5.6 cm), dorsal gland (TDL ≥ 7.3 cm), skin (TDL ≥ 9.2 cm), tactile pelage (TDL ≥ 12.9 cm), coverage pelage (TDL ≥ 17.0 cm), opened eyelids (TDL ≥ 21.5 cm) and tooth eruption (TDL ≥ 24.5 cm). The formula of fetal age was ∛W = 0.079 (t - 27.6), with a linear relationship between TDL and gestational age. The relative weight of tubular gastrointestinal organs, lungs, spleen and thymus increased during fetal development. In contrast, the relative weight of kidneys and liver consistently decreased during the fetal development period. Results of this study indicate the collared peccary is a precocial species and that changes during fetal development are very similar to those in other Suiform species.

Reproductive monitoring of collared peccary females (Pecari tajacu) by analysis of fecal progesterone metabolites.

The measurement of fecal progesterone metabolites (fPM) by enzyme immunoassay analysis is a non-invasive technique that permits gathering reproductive information from wildlife without the stress associated with restraint. In the collared peccary (Pecari tajacu), a high correlation between serum progesterone and fPM levels (r2 = 0.783) suggests that fPM can be used to monitor their reproductive function. We monitored fPM during the estrous cycle of 15 collared peccary females. Estrous cycles averaged 27.9 ±â€¯4.5 days (n = 28), ranging from 21 to 36 days. The luteal phase was 22.2 ±â€¯4.8 days and the inter-luteal phase was 4.3 ±â€¯1.4 days. Mean concentration of fPM across pregnancy were not different from those found during the luteal phase (1230 ±â€¯718 and 1265 ±â€¯584 ng/100 mg dried feces, respectively), however, significant differences were found when luteal phase concentrations were compared only against fPM concentrations during late pregnancy. In addition, late pregnancy fPM concentrations (1893 ±â€¯551 ng/100 mg) were also significantly higher than those in the early (639 ±â€¯339 ng/100 mg) and mid (1134 ±â€¯449 ng/100 mg) pregnancy. For females during the early post-partum period, fPM concentrations were significantly increased (243 ±â€¯118 ng/100 mg) than those of non-cycling females (103 ±â€¯89 ng/100 mg). The analysis of fPM is a simple, non-invasive methodology to detect the ovarian activity in the collared peccary; moreover, it provides a husbandry tool, which may be used to help understand how social structure may impact reproduction.

Postnatal testis development in the collared peccary (Tayassu tajacu), with emphasis on spermatogonial stem cells markers and niche.

Collared peccaries (Tayassu tajacu) present a unique testis cytoarchitecture, where Leydig cells (LC) are mainly located in cords around the seminiferous tubules (ST) lobes. This peculiar arrangement is very useful to better investigate and understand the role of LC in spermatogonial stem cells (SSCs) biology and niche. Recent studies from our laboratory using adult peccaries have shown that the undifferentiated type A spermatogonia (Aund or SSCs) are preferentially located in ST regions adjacent to the intertubular compartment without LC. Following these studies, our aims were to investigate the collared peccary postnatal testis development, from birth to adulthood, with emphasis on the establishment of LC cytoarchitecture and the SSCs niche. Our findings demonstrated that the unique LC cytoarchitecture is already present in the neonate peccary's testis, indicating that this arrangement is established during fetal development. Based on the most advanced germ cell type present at each time period evaluated, puberty (the first sperm release in the ST lumen) in this species was reached at around one year of age, being preceded by high levels of estradiol and testosterone and the end of Sertoli cell proliferation. Almost all gonocytes and SSCs expressed Nanos1, Nanos2 and GFRA1. The analysis of SSCs preferential location indicated that the establishment of SSCs niche is coincident with the occurrence of puberty. Taken together, our findings reinforced and extended the importance of the collared peccary as an animal model to investigate testis function in mammals, particularly the aspects related to testis organogenesis and the SSCs biology and niche.

NEOSPORA CANINUM-SPECIFIC ANTIBODIES IN FREE-RANGING WHITE-LIPPED PECCARIES ( TAYASSU PECARI) FROM THE PERUVIAN AMAZON: DETECTION OF ANTIBODIES IN SERUM AND EVALUATION OF INDIRECT FLUORESCENT ANTIBODY TEST WITH HETEROLOGOUS SECONDARY ANTIBODY.

Neospora caninum is a coccidian parasite originally reported in dogs and widely prevalent in numerous species of wild and domestic animals and has as definitive hosts some species of canids. The white-lipped peccary (WLP) ( Tayassu pecari) is a Tayassuidae mammal, found from Mexico to south of Brazil and north of Argentina. It is a game species with great economic importance in the Peruvian Amazon. Blood samples from 101 WLPs were collected from near or within three different conservation reserves located in the southeastern region of the Peruvian Amazon. For the detection of antibodies against N. caninum, indirect fluorescent antibody tests (IFAT) were performed using collared peccary ( Pecari tajacu) and swine ( Sus scrofa domesticus) heterologous secondary antibodies. For both IFAT tests, the cutoff was 1:50. Positive samples were titrated by a two fold serial dilution. In addition to IFAT, samples were also analyzed using an immunoblotting test (IB) with anti-swine conjugate. To confirm the viability of the anti-swine conjugate, the results of these samples previously tested by a modified agglutination test (MAT) for Toxoplasma gondii were used as reference. From the total of 101 samples tested, 5 (4.9%) were N. caninum positive by the three tests and an extra sample was positive by both IFATs and negative in the IB. Comparing both IFATs and considering IB as the gold standard, the relative sensitivity of IFATs was 100%, the specificity was 98.9%, the positive predictive value was 83.3%, and the negative predictive value was 100%. The agreement between tests was characterized by a κ value of 0.904 (95% confidence interval, 0.717 to 1.0) and an SE of 0.095. This is the first report of N. caninum antibodies in free-ranging T. pecari, and swine and collared peccary conjugate can be used as a secondary antibody for detection of antibodies in Tayassu species.

In vitro maturation of collared peccary (Pecari tajacu) oocytes after different incubation times / Maturação in vitro de oócitos de cateto (Pecari tajacu) após diferentes períodos de incubação

Oocyte in vitro maturation (IVM) is the first step of the in vitro reproductive technologies that enables mature oocytes to be generated ex vivo and after used for embryo production. In this sense, the establishment of culture environment, as oocyte incubation time, is essential for the success of the IVM. Therefore, the study was carried out to investigate the relationship between the meiotic potential and the IVM times of collared peccary oocytes, wild mammals of great commercial and ecological interest. Thus, ovaries were collected of females derived from captivity and transported to the laboratory within 1 hour of slaughtering. The oocytes derived from follicles (3-6mm in diameter) were recovered by aspirated and sliced. Good quality oocytes (evenly granulated cytoplasm with a least one layer of surrounding cumulus cells) were selected and subjected to culture in TCM 199 supplemented with 10µg/mL FSH, 10% FBS and 100µM cysteamine at 38.5°C, 5% CO2 and maximum humidity for 24 or 48 hours. After the incubation period, the nuclear status, the presence of first polar body and the expansion of cumulus cells of oocytes were assessed. The data obtained were analyzed by Fisher exact test (P<0.05). A total of four sessions (2-3 females per session) were performed, resulting in eighteen aspirated and sliced ovaries with normal morphological characteristics. An oocyte recovery rate of about 83.1% (59/71) was obtained with 3.3 oocytes/ovary and 2.3 viable oocytes/ovary. After different incubation times, differences (P<0.05) were observed in 24 and 48 hours for expansion of the cumulus cells (38.1% vs. 100%), presence of first polar body (52.4% vs. 90.5%) and nuclear status in second metaphase (19.0% vs. 76.2%), respectively. In conclusion, 48 hours is suitable time for the in vitro maturation of oocytes derived from collared peccaries when compared to the time of 24 hours, according to the meiotic potential observed. Additional studies should be conducted to improve the quality of the oocyte culture environment, as medium composition, aiming to obtain viable mature oocytes for other in vitro biotechnologies.(AU)

A maturação in vitro (MIV) oocitária é a primeira etapa das tecnologias reprodutivas in vitro que permite que oócitos maturados sejam gerados ex vivo e depois usados para a produção de embriões. Nesse sentido, o estabelecimento do ambiente de cultivo, como o período de incubação de oócitos, é essencial para o sucesso da MIV. Portanto, o estudo foi realizado para investigar a relação entre o potencial meiótico e os períodos de MIV de oócitos derivados de catetos, mamíferos silvestres de grande interesse comercial e ecológico. Para tanto, os ovários foram coletados de fêmeas derivadas de cativeiro e transportados ao laboratório dentro de 1 h após o abate. Os oócitos derivados de folículos (3-6mm de diâmetro) foram recuperados por aspiração e fatiados. Oócitos de boa qualidade (citoplasma uniformemente granulado com pelo menos uma camada circundante de células cumulus) foram selecionados e submetidos ao cultivo em TCM 199 suplementado com 10µg/mL de FSH, 10% de SFB e 100μM de cisteamina a 38,5°C, 5% de CO2 e umidade máxima por 24 e 48 h. Após o período de incubação, o estado nuclear, a presença do primeiro corpúsculo polar e a expansão das células do cumulus dos oócitos foi avaliada. Os dados obtidos foram analisados pelo teste exato de Fisher (P<0,05). Um total de quatro sessões (2-3 fêmeas por sessão) foi realizado, resultando em dezoito ovários aspirados e fatiados com características morfológicas normais. Uma taxa de recuperação oocitária de aproximadamente 83,1% (59/71) foi obtida com 3,3 oócitos/ovário e 2,3 oócitos viáveis/ovário. Após diferentes períodos de incubação, diferenças (P<0,05) foram observadas entre 24 e 48 h para a expansão das células cumulus (38,1% vs. 100%), presença de primeiro corpúsculo polar (52,4% vs. 90,5%) e estado nuclear na segunda metáfase (19,0% vs. 76,2%), respectivamente. Em conclusão, 48 h é o período adequado para a maturação in vitro de oócitos derivados de catetos quando comparado ao tempo de 24 h, de acordo com o potencial meiótico observado. Estudos adicionais devem ser conduzidos para melhorar a qualidade do ambiente de cultivo oocitário, como a composição de meio, objetivando obter oócitos maturados viáveis para outras biotecnologias in vitro.(AU)

Nutrition of Six Selected Neo-Tropical Mammals in Trinidad and Tobago with the Potential for Domestication.

This review highlights the available literature on the nutrition of six neo-tropical animals with the potential for domestication-the agouti (Dasyprocta leporina/D. aguti), lappe (Agouti paca), capybara (Hydrochoerus hydrochaeris), manicou/opossum (Didelphis marsupialis insularis), collared peccary (Peccary tajucu) and the red brokcet deer (Mazama americana). Over 100 references were used, spanning over 100 years. The earliest being 1915 and the most recent being 2018. The references used in this review were synthesized to give a detailed look of the dentition, anatomy of the gastrointestinal tract and type of feed these animals consume. Nutritional requirements of the animals are required to understand what is needed for growth, maintenance and reproduction of each physiological stage. The agouti (D. leporina/D. aguti) was observed to be a monogastric mammal that fed primarily on fruits, seeds, animal matter and practiced caecotrophy. The lappe/paca (C. paca/A. paca) was described as a strict herbivore and a frugivore which practiced caecotrophy, with a diet that varied throughout the year, according to food availability. The capybara (H. hydrochaeris) was found to be the largest known rodent and was described as a semiaquatic hindgut fermenter that practiced caecotrophy. The manicou/opossum (D. marsupialis insularis) was found to be an omnivore with a simple stomach. The collared peccary (T. tajacu) was found to be frugivorous. Their unique stomach enabled them to consume a wide variety of feedstuff, allowing them to be found in a wide range of habitats. The red brocket deer (M. americana), a ruminant, was described as a browser that consumed mainly fruits and seeds and they frequented mineral lick. Knowledge of what they consume in the wild is important, so that we know what to feed in captivity. There is also the need to evaluate captive diets while trying to domesticate these mammals and develop nutrient requirement tables for these neo-tropical animals. Finally, an understanding of the dentition and gastrointestinal tract is important to increase efficiency (nutritional and cost). These six neo-tropical mammals were chosen due to their prevalence as game species in Trinidad and Tobago.

Identification of Mycobacterium species and Rhodococcus equi in peccary lymph nodes.

Mycobacterium species and the virulence-associated proteins (vapA, vapB, and vapN genes) of Rhodococcus equi isolated from 330 lymph nodes of collared peccaries (Tayassu tajacu) and white-lipped peccaries (Tayassu pecari) intended for human consumption were investigated. Thirty-six (10.9%) R. equi strains were isolated; 3.3% (n = 11/330) were from white-lipped peccary lymph nodes, and 7.6% (25/330) were from collared peccary lymph nodes. Among the 11 isolates of R. equi from the white-lipped peccaries, 90.9% (n = 10/11) were obtained from the mesenteric lymph nodes, and only 9.1% (n = 1/10) were obtained from the mediastinal lymph nodes. In the 25 isolates of R. equi obtained from the collared peccaries, 40.0% (n = 10/25) were recovered from the mesenteric lymph nodes, 36% (n = 9/25) from the submandibular lymph nodes, and 24.0% (n = 6/25) from the mediastinal lymph nodes. No vapA, vapB, or vapN genes (plasmidless) or three host-associated types (pVAPA, pVAPB, and pVAPN) were identified among the R. equi isolates. Mycobacterium species were isolated in 3.03% (n = 10/330) of all the lymph nodes analyzed. Among the 10 mycobacterial isolates, 60% (n = 6/10) were from the white-lipped peccary lymph nodes, and 40% (n = 4/10) were from the collared peccary lymph nodes. Ten Mycobacterium species were detected by PCR-PRA with a predominance of M. avium type 1. Sequencing of the hsp65 and rpob genes revealed mycobacteria that were saprophytic (M. sinense and M. kumamotonense) and potentially pathogenic (M. colombiense and M. intracellulare) to humans and animals. To our knowledge, this is the first description of R. equi and/or mycobacterial species identified in the lymph nodes of peccary specimens. R. equi (plasmidless) and the mycobacterial species described here have been reported as causes of pulmonary and extrapulmonary infections in both immunocompetent and immunocompromised humans.

Combination of ethylene glycol with sucrose increases survival rate after vitrification of somatic tissue of collared peccaries (Pecari tajacu Linnaeus, 1758) / Combinação de etilenoglicol com sacarose aumenta a taxa de sobrevivência após a vitrificação de tecido somático de catetos (Pecari tajacu Linnaeus, 1758)

The cryopreservation of somatic tissue in collared peccaries promotes an alternative source of genetic material of this specie. The solid-surface vitrification (SSV) is a great option for tissue conservation; nevertheless, the optimization of SSV requirements is necessary, especially when referred to cryoprotectants that will compose the vitrification solution. Therefore, the aim was to evaluate the effect of the presence of 0.25 M sucrose in addition to different combinations (only or association) and concentrations (1.5 M or 3.0 M) of ethylene glycol (EG) and/or dimethyl sulfoxide (DMSO) in the somatic tissue vitrification of collared peccaries. Subsequently, we tested six combinations of cryoprotectants with or without sucrose in Dulbecco modified Eagle medium (DMEM) plus 10% fetal bovine serum (FBS). Thus, 3.0 M EG with sucrose was able to maintain normal tissue characteristics compared with non-vitrified (control), especially for the volumetric ratio of epidermis (61.2 vs. 58.7%) and dermis (34.5 vs. 36.6%), number of fibroblast (90.3 vs. 127.0), argyrophilic nucleolar organizer region (AgNOR) ratio (0.09 vs. 0.17%) and nucleus area (15.4 vs. 14.5 µm2) respectively. In conclusion, 3.0 M EG with 0.25 M sucrose and 10% FBS resulted in a better cryoprotectant composition in the SSV for somatic tissue of collared peccaries.(AU)

A criopreservação de tecido somático em catetos promove uma fonte alternativa de material genético nesta espécie. A vitrificação em superfície sólida (VSS) é uma ótima opção para a conservação do tecido; contudo, a otimização dos requerimentos da VSS é necessária, especialmente quanto aos crioprotetores que irão compor a solução de vitrificação. Portanto, o objetivo foi avaliar o efeito da presença de 0,25 M de sacarose em adição com diferentes combinações (individual ou associação) e concentrações (1,5 M ou 3,0 M) de etilenoglicol (EG) e/ou dimetilsulfóxido (DMSO) na vitrificação de tecido somático de catetos. Subsequentemente, nós testamos seis combinações de crioprotetores com ou sem sacarose em meio de Eagle modificado por Dulbecco (DMEM) acrescido de 10% de soro fetal bovino (SFB). Assim, 3,0 M de EG com sacarose foi capaz de manter as características normais do tecido comparado com o não vitrificado (controle), especialmente para a proporção volumétrica da epiderme (61,2 vs. 58,7%) e derme (34,5 vs. 36,6%), número de fibroblastos (90,3 vs. 127,0), razão da região argirófila organizadora de nucléolo (AgNOR) (0,09 vs. 0,17%) e área do núcleo (15,4vs.14,5 µm2), respectivamente. Em conclusão, 3,0 M de EG com 0,25 M de sacarose e 10% de SFB resultaram na melhor composição de crioprotetores na VSS para tecido somático de catetos.(AU)

Progesterone and estradiol profiles in different reproductive stages of captive collared peccary (Pecari tajacu) females assessed by fecal metabolites.

The study determined the fecal progesterone and estradiol profiles in different reproductive stages of captive collared peccary (Pecari tajacu) females from eastern Mexico. Fifteen adult females were included. At the start of the study the females were either pregnant (early, mid, or late pregnancy), lactating, or non-lactating of unknown pregnancy status. Feces from each female were collected once a week during nine consecutive months to determine concentrations of fecal progesterone and estradiol metabolites using ELISA. Progesterone was similar in early (2048±285ng/g), mid (2254±274ng/g), and late pregnancy (2491±374ng/g), and in early-pregnant and non-lactating females (1154±274ng/g). Progesterone in lactating females (442±255ng/g) was lower than in females at any stage of pregnancy, but was similar to non-lactating females. Overall progesterone in pregnant females (2229±173ng/g) was higher than in lactating and non-lactating females together (772±189ng/g). Estradiol was similar in early (66±8ng/g), mid (83±9ng/g), late pregnant (109±15ng/g), and non-lactating females (64±9ng/g). Estradiol in lactating females (34±8ng/g) was similar to estradiol in early-pregnant and non-lactating females, but was lower than in females in late and mid pregnancy. Overall estradiol in pregnant females (79±6ng/g) was similar to non-lactating females, but higher than in lactating females. The progesterone and estradiol profiles of captive collared peccary females at different reproductive stages were determined by assessing concentrations of fecal hormone metabolites.

In vitro culture of somatic cells derived from ear tissue of collared peccary (Pecari tajacu Linnaeus, 1758) in medium with different requirements / Cultivo in vitro de células somáticas derivadas de tecido auricular de cateto (Pecari tajacu Linnaeus, 1758) em meio com diferentes requerimentos

The maintenance of metabolic activities during the in vitro culture of somatic cells of wild animals, especially collared peccary (Pecari tajacu), is an interesting step in conservation of these cells for the use in nuclear transfer. In this context, it is necessary to optimize the culture conditions of somatic cells by the establishment of appropriate supplementation to the media. Therefore, this study aimed to analyze the composition of the culture means of somatic cell derived from ear tissue of collared peccaries, evaluating concentrations of fetal bovine serum (FBS; 10% vs. 20%) and epidermal growth factor (EGF; 5ng/mL vs. 10ng/mL). Tissues were submitted to primary culture and subcultures for 40 days and cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity to develop the growth curve and to determine the population doubling time (PDT), viability, and functional/metabolic activity. No difference was observed between the concentrations of FBS for several parameters, except for viability [FBS10: 85.6% vs. FBS20: 98.2%], PDT [FBS10: 155.4h vs. 77.2h], and functional/metabolic assay [FBS10: 0.57-0.55 vs. FBS20: 0.82-0.99 (D5-D7)]. For the EGF in culture, no difference was observed in the evaluated parameters. In all experiments, the growth curves were typical S-shape and the cells passed through a lag, logarithmic, and plateau phase. In conclusion, 20% FBS is suitable for the recovery of somatic cells; nevertheless, EGF does not improve the quality of growing these cells. To our knowledge, this is the first study culturing somatic cells of collared peccaries.(AU)

A manutenção das atividades metabólicas durante o cultivo in vitro de células somáticas de animais silvestres, especialmente cateto (Pecari tajacu), é uma etapa interessante na conservação dessas células para o uso na transferência nuclear. Nesse contexto, é necessário aperfeiçoar as condições de cultivo de células somáticas pelo estabelecimento de suplementações apropriadas aos meios. Portanto, este estudo objetivou analisar a composição dos meios de cultivo de células somáticas derivadas de tecido auricular de catetos, avaliando concentrações de soro fetal bovino (SFB; 10% vs. 20%) e fator de crescimento epidermal (EGF; 5 ng/mL vs. 10 ng/mL). Para tanto, tecidos foram submetidos ao cultivo primário e subcultivos por 40 dias e células foram analisadas por morfologia, adesão, subconfluência, e atividade proliferativa pelo desenvolvimento da curva de crescimento e determinação do time de duplicação da população (PDT), viabilidade, e atividade funcional/metabólica. Nenhuma diferença foi observada entre as concentrações de SFB para os vários parâmetros, exceto para viabilidade [SFB10: 85,6% vs. SFB20: 98,2%], PDT [SFB10: 155,4 h vs. 77,2 h], e atividade funcional/metabólica [SFB10: 0,57-0,55 vs. SFB20: 0,82-0,99 (D5-D7)]. Para o EGF em cultivo, nenhuma diferença foi observada nos parâmetros avaliados. Em todos os experimentos, as curvas de crescimento foram típicas de forma S e as células passaram por uma fase lag, logarítmica e platô. Em conclusão, 20% de SFB é adequado para a recuperação de células somáticas; contudo, EGF não melhora a qualidade de crescimento dessas células. Ao nosso conhecimento, este é o primeiro estudo cultivando células somáticas de catetos.(AU)

Sexual behavior of Pecari tajacu (Cetartiodactyla: Tayassuidae) during periovulatory and early gestation periods.

The goal of this study was to describe the sexual behavior in female and male collared peccary. Twenty females and twelve males were monitored in familiar and non-familiar units for two 60 days periods. During both phases, we recorded 2747 sexual interactions initiated by 20 different females toward males and 4461 sexual interactions initiated by 12 males toward females. The frequency of sexual interactions initiated per female significantly increased from proestrus to estrus, and they were significantly more frequently courted. Females initiated olfactory inspections 15.42 times more and were mounted 22.6 times more during estrus than during proestrus. Nulliparous and primiparous females copulated only when exposed to non-parental males. After estrus, the frequency of sexual interactions received by females sharply decreased. One mating event was recorded during the first gestation week and 31 mountings were observed after the second week. In conclusion, the behavioral monitoring is a useful procedure for the recognition of estrus. Our results suggest that ovulation may be associated with the end of the estrus, which will support future work in assisted reproduction in this species. To promote good handling practices, females of reproductive age should be removed from their family unit of origin.

Extramedullary plasmacytoma in a captive collared peccary (Pecari tajacu) / Plasmocitoma extramedular em cateto criado em cativeiro (Pecari tajacu)

An extramedullary plasmacytoma case in a captive collared peccary (Pecari tajacu) is reported. The animal, a female aging three years old, had a medical history of diffusely distributed skin and mucocutaneous junction lesions, associated with swollen lymph nodes. Clinical examination and complementary exams (complete blood count, biochemical analysis, skin scraping to search mites and fungal culture) were performed. Thirty days after examination, the animal died. At necropsy, multiple consistent nodules, aseptic pustules and swollen lymph nodes were found. On histological exams of the skin and oral mucosa, we observed a large number of round cells forming masses organized in nests, sheets, and cords of cells in a well-vascularized fibrovascular tissue. Neoplastic plasma cells infiltrated between the fibers and the lamina propria of smooth muscle. Spaces among the cell masses were filled with some eosinophil and fluid. Most of the cells were well differentiated, presenting a perinuclear clear zone. In some points, the cells were pleomorphic. The plasma cells presented eccentric, basophilic and spherical nuclei, showing a dense to organized chromatin with distinct nucleoli. Binucleate cells were observed, but multinucleated giant cells were rare. Oral mucosa and lymph nodes tested by immunohistochemical analyses were positive for Mb-1, with a multifocal distribution. In regard to Bcl-2, the neoplastic cells were intermittent weakly positive. So, an extramedullary plasmacytoma was diagnosed in the collared peccary considering the location, the histopathological and immunohistochemical findings.(AU)

Um caso de plasmocitoma extramedular em cateto criado em cativeiro (Pecari tajacu) está sendo relatado. O animal, uma fêmea com três anos de idade, apresentou um histórico médico de lesões de pele, envolvendo junção mucocutânea e de aumento de linfonodos. Exame clínico e exames complementares (hemograma completo, análise bioquímica, raspado de pele para pesquisa de ácaros e cultura fúngica) foram realizados. Após 30 dias, o animal morreu. Na necropsia, verificou-se a presença de múltiplos nódulos, pústulas assépticas e aumento dos linfonodos. Na pele e mucosa oral, histologicamente as massas consistiam em ninhos, lençóis e cordões de células redondas, e um estroma fibrovascular bem vascularizado. Os plasmócitos foram observados infiltrados entre as fibras e músculo liso da lâmina própria. Espaços contendo eosinófilos, fluido e células livres estavam presentes na massa. A maioria das células estava bem diferenciada, com uma zona perinuclear clara, mas algumas células demonstraram-se pleomórficas. Os plasmócitos apresentavam núcleo excêntrico, redondo, basófilo, e pontilhado, com cromatina variando de densa a grosseiramente organizada e nucléolos distintos. O citoplasma finamente granular de anfifílico para basofílico. As células foram interpretadas como plasmócitos neoplásicos. Células binucleadas foram observadas e células gigantes multinucleadas eram raras. Na imunohistoquímica de tecidos da mucosa oral e de linfonodos observou-se positividade moderada e multifocal para Mb-1. As células tumorais revelaram positividade fraca e intermitente para Bcl-2. Com base na localização, achados histopatológicos e imuno-histoquímicos, um plasmocitoma extramedular foi diagnosticado.(AU)

Morphology and vascularization of the corpus luteum of peccaries (Pecari tajacu, Linnaeus, 1758) throughout the estrous cycle / Morfologia e vascularização do corpo lúteo de catetos (Pecari tajacu, Linnaeus, 1758) ao longo do ciclo estral

The current paper characterizes the changes in morphology and vascularization of the corpus luteum of collared peccaries during the estrous cycle and correlates progesterone synthesis (P4). Twenty females were subjected to a treatment for estrus synchronization; an ear implant containing 1.5 mg of norgestomet was implanted on D0, whereas on D9 the peccaries received an IM injection of eCG 200UI and 50g of PGF2a. The animals were divided into four groups (G1, G2, G3 and G4) and euthanized on post-ovulation days 3, 12, 18 and 22. The ovaries were collected and the corpora lutea were measured and processed for histological and vascular density (Dv). Blood was collected for dosage of P4 serum. The morphology of the ovaries, the corpora lutea and P4 varied significantly during the estrous cycle (P<0.001). There was a significant co-relationship between weight and length of the ovaries and CL (r = 0.66, r = 0.52, P<0.05, respectively) and between weight, length and width of the CL and P4 (r = 0.51, r = 0.54 and r = 0.68, P<0.05, respectively). The luteal Dv was highly influenced by the estrous cycle phase (P<0.0001). The P4 and luteal Dv concentrations were higher in G2 and evidenced maximum secretory activity, with a highly significant correlation (P<0.0001). Assessed lutein parameters may estimate the phase of the estrous cycle in peccaries and the functional activity of the corpus luteum.

Objetivou-se caracterizar as variações na morfologia e vascularização do corpo lúteo (CL) de catetos durante ciclo estral (CE) e correlacioná-las com a concentração de progesterona (P4). Vinte fêmeas de cateto foram submetidas a tratamento de sincronização do estro; no D0 receberam implante auricular contendo 1,5mg de norgestomet, no D9 injeção via IM de 200UI de eCG e 50µg de PGF2α. Os animais foram divididos em quatro grupos (G1, G2, G3 e G4) e eutanasiados nos dias três, 12, 18 e 22 pós-ovulação. Os ovários foram coletados e os CL foram mensurados e processados para avaliação histológica e da densidade vascular (Dv). O sangue foi coletado para dosagem da P4 sérica. A morfologia dos ovários, CL e a concentração de P4 variaram significativamente durante o CE (P<0,001). Houve correlação significativa entre peso e comprimento dos ovários e CL (r = 0,66, r = 0,52, P<0,05, respectivamente) e entre peso, comprimento e largura do CL e a concentração de P4 (r=0,51, r=0,54 e r=0,68; P<0,05, respectivamente). A Dv do CL se mostrou muito influenciada pela fase do CE (P<0,001) e apresentou alta correlação significativa (P< 0,001). No G2 os maiores valores de P4 e Dv confirmaram máxima atividade secretória do CL nesse estádio. Os parâmetros luteínicos avaliados podem ser usados para estimar a fase do ciclo estral em catetos e a atividade funcional do CL.

Sperm characteristics following freezing in extenders supplemented with whole egg yolk and different concentrations of low-density lipoproteins in the collared peccary (Pecari tajacu).

The aim of the current study was to compare sperm quality characteristics of the collared peccary (Pecari tajacu) following freezing in extenders supplemented with whole egg yolk and different concentrations of low-density lipoproteins (LDL). Semen from 11 adult males was obtained by electroejaculation and evaluated for sperm motility, vigor, morphology as well as membrane integrity analyzed by the hypo-osmotic swelling (HOS) test and a fluorescent staining. Moreover, the semen was diluted in a Tris-based extender containing 20% egg yolk (control group) or 5, 10 or 20% LDL (treatment groups). The semen samples were frozen in liquid nitrogen and thawed in a water bath for 60s at 37°C. The treatments did not affect (p>0.05) sperm vigor, morphology or membrane integrity analyzed by the HOS test. However, post-thaw sperm motility was significantly higher (p<0.05) in the extender supplemented with 20% LDL (36.4 ± 5.3%) compared with the egg yolk extender and extender supplemented with 10% LDL. Furthermore, the percentage of membrane-intact frozen-thawed spermatozoa analyzed by the fluorescent staining was significantly higher (p<0.05) in the extender supplemented with 20% LDL (27.4 ± 6.5%) than in the other groups. In conclusion, 20% LDL can be used to substitute the whole egg yolk as a cryoprotective additive for freezing semen of the collared peccary.

Postmortem findings in collared peccaries raised in captivity in northeastern Brazil / Achados post-mortem em catetos criados em cativeiro no Nordeste do Brasil

This study is a retrospective examination of diseases in collared peccaries that were diagnosed by the Veterinary Pathology Laboratory, Universidade Federal Rural do Semiárido. Necropsy and histological examination were performed from 2005 to 2010. Of the 50 necropsied collared peccaries, 24% died due to restraint and capture myopathy; 18% died from trauma; and the remainder was diagnosed with splenic hemangioma (6%), enterolithiasis (6%), gastritis (6%), gastric ulcer (4%), intestinal volvulus (4%), gastric volvulus (2%), mammary carcinoma (2%), polycystic kidney disease (2%), pyometra (2%), and suppurative bronchopneumonia (2%). Twelve animals remained undiagnosed, seven of which (14%) were in advanced autolytic condition and five of which (10%) had no gross or microscopic lesions that were compatible with disease. This paper describes illnesses that have not been reported in the collared peccary, focusing on their clinical and pathological aspects.(AU)

Este trabalho teve por objetivo realizar um estudo retrospectivo sobre as doenças de catetos diagnosticadas pelo Laboratório de Patologia Veterinária da Universidade Federal Rural do Semi-Árido, através do exame de necropsia e histológico no período de 2005 a 2010. Dos 50 catetos submetidos à necropsia, as percentagens das enfermidades diagnosticadas foram: miopatia por captura e contenção (24%), morte por traumas (18%), hemangioma do baço (6%), enterolitíase (6%), gastrite (6%), úlcera gástrica (4%), vólvulo intestinal (4%), vólvulo gástrico (2%), carcinoma mamário (2%), doença renal policística (2%), piometra (2%) e broncopneumonia supurativa (2%). Onze (22%) animais permaneceram sem diagnóstico, dos quais sete (10%) apresentavam avançado estado autolítico e cinco (14%) não foram observadas lesões macroscópicas e microscópicas compatíveis com nenhuma enfermidade. Este estudo apresenta relatos de doenças ainda não descritas em catetos, com enfoque nos aspectos clínicos e patológicos.(AU)

Relações entre a câmara de Neubauer a espectrofotometria utilizadas para a determinação da concentração espermática de catetos (Pecari tajacu) / Relationship between Neubauer counting chamber and spectrophotometer used for the sperm concentration determination in collared peccaries (Pecari tajacu)

Em ejaculados provenientes de 28 catetos, verificou-se a existência de relações entre a concentração espermática determinada por meio da câmara de Neubauer e a tramitância observada por espectrofotometria, utilizando comprimentos de onda variando de 530 a 590nm. Os ejaculados apresentaram uma concentração média de 283,9±30,8x106 espermatozoides mL-1, com variação de 30 a 640x106 espermatozoides mL-1. Os valores para tramitância variaram entre 36,9 a 96,3, nos diferentes comprimentos de onda. Não foram detectadas relações significativas entre os dois métodos (P>0,05). Dessa forma, não se recomenda a espectrofotometria para os procedimentos de rotina na determinação da concentração espermática em catetos.

In ejaculates derived from 28 collared peccaries, we verified the existence of relationships between sperm concentration determined by the Neubauer counting chamber and the tramitance verified by spectrophotometer, under wavelengths varying from 530 to 590nm. Ejaculates presented a concentration of 283.9±30.8x106sperm m-1, varying from 30 to 640x106sperm mL-1. Values for tramitance varied from 36.9 to 96.3, under different wavelengths. No significant relationship was verified between two methods (P>0.05). Thus, the spectrophotometer is not recommended for routine procedures of sperm concentration measurement in collared peccaries.

Determinação da concentração alveolar mínima do isofluorano em catetos (Tayassu tajacu) / Isoflurane minimum alveolar concentration in collared peccary (Tayassu tajacu)

A anestesia inalatória vem sendo amplamente difundida na medicina veterinária, no entanto seu uso em animais selvagens ainda é restrito, não sendo observado nenhum estudo referente à sua utilização na espécie Tayassu tajacu. O objetivo da pesquisa foi determinar a concentração alveolar mínima (CAM) do isofluorano em catetos e apresentar os efeitos desta administração sobre as variáveis hemodinâmicas e respiratórias, como também a qualidade da recuperação anestésica. Utilizou-se 10 animais, machos, com idade variando de 1 a 3 anos oriundos do Centro de Multiplicação de Animais Silvestres da Universidade Federal Rural do Semi-Árido, Brasil. Todos os animais tiveram anestesia induzida com 7mg.kg-1 de propofol e posteriormente foram conectados a circuito anestésico com isofluorano e oxigênio 100 por cento. O estímulo noceptivo supramáximo adotado foi pinçamento interdigital, o qual era realizado após 15 minutos de espera para cada concentração de isofluorano fornecida. Ao ser observada resposta negativa frente ao estímulo a concentração era reduzida em 20 por cento, quando verificada resposta positiva o estímulo era cessado, calculando-se a partir daí o valor da CAM. [...] A recuperação anestésica foi tranquila e rápida. Concluiu-se que a CAM do isofluorano para catetos foi maior que a observada em espécies afins. O isofluorano pode ser utilizado nesta espécie, sendo considerado seguro e eficaz. A recuperação dos animais após anestesia com isofluorano foi livre de excitação.

Inhalation anesthesia has been widespread in veterinary medicine. Nevertheless, its use in wild animals is still limited, having no studies on its use been observed in the species. The objective of the research was to determine the isoflurane minimum alveolar concentration (MAC) in peccaries and present the effects of its administration on the hemodynamic and respiratory variables, as well as data concerning the anesthesia recovery. The study used 10 male animals with age ranging from one to three years, from the Centro de Multiplicação de Animais Silvestres of Universidade Federal Rural do Semi-Árido, Brazil. All the animals had anesthesia induced with propofol 7mg.kg-¹, were intubated and connected to the anesthetic circuit with isoflurane and 100 percent oxygen. The supramaximal noxious stimulation used was the interdigital pinch, which was performed after 15 minutes of waiting for each provided isoflurane concentration. When negative response to the stimulus was observed, the concentration was reduced by 20 percent; when positive response was verified, the stimulus was stopped, being the CAM value calculated from that point. [...] Recovery was quiet and smooth. It was concluded that the isoflurane MAC for peccaries was greater than that observed in related species. Isoflurane can be used in this species, being considered safe and effective. The animals' recovery after anesthesia with isoflurane was free from excitement.

Influence of recovery method and centrifugation on epididymal sperm from collared peccaries (Pecari tajacu Linnaeus, 1758).

In order to establish protocols for gamete recovery from accidentally killed wild animals, or to take advantage of those slaughtered by captive breeders, we assess the influence of two methods on the recovery of epididymal sperm from collared peccaries, and verify the effect of centrifugation on such gametes. Genitalia from nine animals were used. For each animal, one epididymis was processed by flotation and the other was processed by retrograde flushing, both using a buffered media based on Tris. Following recovery, sperm were evaluated for motility, vigor, viability, functional membrane integrity, and morphology. A 1-mL aliquot of each sample was centrifuged, the supernatant removed, and the pellet suspended and evaluated as fresh samples. The sperm characteristics did not differ between the samples collected by flotation or retrograde flushing (P < 0.05). Centrifugation promoted an increase in head and tail defects, thus reducing the percentage of viable sperm (P < 0.05). No other parameter assessed for both methods was affected by centrifugation. In conclusion, epididymal sperm from collared peccaries can be efficiently collected through flotation or retrograde flushing, but not when either is followed by centrifugation.

Ovarian folliculogenesis in collared peccary: Pecari tajacu (Artiodactyla: Tayassuidae)

The sustainability and production of collared peccary (Pecari tajacu) has been studied in the last few years; however, further information on its reproduction is necessary for breeding systems success. Understanding folliculogenesis aspects will contribute to effective reproductive biotechniques, which are useful in the preservation and production of wildlife. The aim of this study was to evaluate the ovarian folliculogenesis in collared peccary. Ovaries from six adult females of collared peccary were obtained through ovariectomy and analyzed. These were fixed in aqueous Bouin’s solution and sectioned into 7μm slices, stained with hematoxilin-eosin and analyzed by light microscopy. The number of pre-antral and antral follicles per ovary was estimated using the Fractionator Method. The follicles, oocytes and oocyte nuclei were measured using an ocular micrometer. Results showed that the length, width, thickness, weight, and the gross anatomy of the right and left ovaries were not significantly different. However, the mean number of corpora lutea was different between the phases of the estrous cycle (p<0.05), with the highest mean in the luteal phase. Primordial follicles were found in the cortex; the oocytes were enveloped by a single layer of flattened follicular cells. In the primary follicles, proliferation of the follicular cells gave rise to cuboidal cells (granulosa cells). The secondary follicle was characterized by two or more concentric layers of cuboidal cells (granulosa), beginning of antrum formation, and the presence of pellucid zone and theca cells. Antral follicles were characterized by a central cavity (antrum), the presence of cumulus oophorus and theca layers (interna and externa). In the right ovary, the values of the primordial and primary follicles were similar, but significantly different from the secondary ones (p<0.05). In the left ovary, significant differences were observed between all follicles in the follicular phase (p<0.05); the mean number of primordial and primary follicles was similar in the luteal phase. The mean number of pre-antral follicles and antral follicles in the follicular phase was higher in the left ovary (p<0.05). The mean number of antral follicles in the luteal phase was similar in both ovaries. We also found significant differences in mean diameter of preantral follicles, oocyte, granulosa layer and oocyte nucleus during the estrous cycle. In the antral follicles a significant difference was observed only in follicular diameter (p<0.05). The predominance of active primordial and primary follicles was found in both phases; otherwise the secondary follicles and antral follicles showed a high degree of degeneration. The results obtained in the present work will strengthen the development of biotechnology programs to improve the productive potential and conservation of the collared peccary.

La sustentabilidad y la producción de pecarí de collar (Pecari tajacu) han sido estudiados en los últimos años, sin embargo, más información sobre su reproducción es necesaria para el éxito de los sistemas de crianza . La comprensión de los aspectos relacionados con la foliculogénesis contribuirá con la aplicación de biotécnicas de reproducción, las cuales son útiles en la preservación y la producción de la vida silvestre. El objetivo de este estudio fue obtener datos sobre la población folicular del ovario de pecarí de collar. En relación con la población folicular en el ovario derecho, los valores de los folículos primordiales y primarios fueron similares, pero se observó que había una diferencia significativa (p<0.05) con el secundario. En el ovario izquierdo, la fase folicular presentó diferencias significativas (p<0.05) entre todos los folículos, y en la fase lútea el número medio de folículos primordiales y primarios fueron similares. Ahora bien, con respecto a la población de folículos antrales, en la fase folicular, se observaron diferencias significativas entre los ovarios (p<0.05), y de forma similar en la fase lútea. También se encontraron diferencias significativas en el diámetro medio de los folículos preantrales, oocitos, la capa granulosa y el núcleo del oocito durante las fases del ciclo estral. Asimismo, en los folículos antrales se observó diferencia estadísticamente significativa (p<0.05) en el diámetro folicular. En ambas fases del ciclo estral, se encontró el predominio de folículos primordiales y primarios en desarrollo, por otro lado, los folículos secundarios y los folículos antrales mostraron un alto grado de degeneración. Los resultados aquí presentes son necesarios para el desarrollo de los programas de mejoramiento y conservación.