Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology.

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox = 46, crab-eating fox = 55) and feces (pampas fox = 84, crab-eating fox = 2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan™ probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n = 1), thus it was also detected from pampas fox tissues (n = 1). Meanwhile, T. gondii was found in tissues of pampas (n = 1) and crab-eating (n = 1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife.

First assessment of hoary fox (Lycalopex vetulus) seasonal ovarian cyclicity by non-invasive hormonal monitoring technique.

Reproduction is key to species survival, and reproductive physiology represents a high priority investigative area for conservation biology, as it provides a basic understanding of critical life-history traits, information that is helpful for the establishment of management strategies. Here, we generated knowledge about the reproductive endocrinology of the hoary fox (Lycalopex vetulus), a small canid (2.5-4 kg) endemic to open areas of the Brazilian Cerrado and listed in the Brazilian National List of Endangered species. Specifically, we utilized non-invasive hormone monitoring methods to assess oestrogen and progestagen metabolites from eight female hoary foxes housed in five zoological institutions in the state of São Paulo-Brazil. We observed the elevations of oestrogen and progestagen metabolites between July and September in six of the eight females. No significant evidence of ovarian activity was observed during other months. Two females, who shared the same enclosure, did not show a pattern of reproductive cyclicity. Based on these characteristics, we concluded that captive hoary foxes are seasonal monoestric, with the beginning of the oestrus cycle occurring mainly in July followed by 2 months of the luteal phase when conception does not occur. We suggest the dosage of faecal metabolites of estradiol and progesterone could be used to differentiate the reproductive period from a non-reproductive period in Lycalopex vetulus females, providing relevant information about their reproductive biology that may contribute to species conservation and management strategies, such as increased ex situ reproductive success.

Dimensions and skeletopy of kidneys in two populations of Cerdocyon thous (Linnaeus, 1766) / Dimensões e esqueletopia dos rins em duas populações de Cerdocyon thous(Linnaeus, 1766)

Cerdocyon thous is the South American canid with great geographic distribution. To the south of Ecuador, two isolated populations have been identified, living in different average temperatures and food availability. The objective was to measure the length, width, thickness and volume of the kidneys, length of renal vessels, and verify the renal skeletopy in two populations of C. thous. Kidneys and renal vessels were measured from 34 cadavers collected on highways in the Brazilian territory. From the Atlantic Forest biome (latitude 22º), 14 specimens (seven males and seven female) were analyzed, and from the Pampa biome (latitude 29º), 20 specimens (eight males and twelve female). On average, in the right antimere the kidneys measured 49.9 ´ 25.2 ´ 24.4 mm, had a volume of 16.5 cm3, the renal artery measured 21.3 mm and the renal vein 19.4 mm. In the left antimere, the kidneys measured 49.3 ´ 24.4 ´ 22.8 mm, with a  volume of 14.6 cm3 and the artery and vein measured 21.0 mm and 28.4 mm, respectively. The right kidney was always cranial and predominantly positioned ventrally to vertebrae L1L3, while the left one was positioned ventrally to vertebrae L2L4. There was no difference in the comparison between sexes or antimeres. Most renal dimensions were significantly higher in the specimens from the Pampa biome, possibly due to the body size and type of diet.

Cerdocyon thousé o canídeo sul americano com maior distribuição geográfica. Ao sul do Equador, duas populações distintas são reconhecidas, submetidas a diferentes temperaturas médias e disponibilidade de alimento. Objetivou-se mensurar o comprimento, largura, espessura e volume dos rins, comprimento dos vasos renais e esclarecer a esqueletopia renal em duas populações isoladas de C. thous.Para tal, foram estudados rins e vasos renais de 34 cadáveres coletados mortos em rodovias brasileiras. Catorze espécimes (sete machos e sete fêmeas) são provenientes do bioma Mata Atlântica (latitude 22º) e 20 espécimes (oito machos e doze fêmeas) do bioma Pampa (latitude 29º). Na média, os rins direitos mediram 49,9x25,2x24,4 mm, tinham um volume de 16,5 cm3e a artéria renal direita mediu 21,3 mm e a veia renal direita 19,4 mm. Os rins esquerdos mediram 49,3x24,4x22,8 mm tinham volume de 14,6 cm3e as artérias e veias renais direitas mediram 21,0 mm e 28,4 mm, respectivamente. Os rins direitos foram sempre craniais e predominantemente posicionados ventralmente às vertebras L1 a L3, enquanto os rins esquerdos estavam posicionados majoritariamente ao nível de L2 a L4. Não houve diferença na comparação das medidas entre os sexos ou antímeros. Praticamente todasas medidas renais foram significativamente maiores nos espécimes do bioma Pampa, possivelmente devido

Dimensions and skeletopy of kidneys in two populations of Cerdocyon thous (Linnaeus, 1766) / Dimensões e esqueletopia dos rins em duas populações de Cerdocyon thous(Linnaeus, 1766)

Cerdocyon thous is the South American canid with great geographic distribution. To the south of Ecuador, two isolated populations have been identified, living in different average temperatures and food availability. The objective was to measure the length, width, thickness and volume of the kidneys, length of renal vessels, and verify the renal skeletopy in two populations of C. thous. Kidneys and renal vessels were measured from 34 cadavers collected on highways in the Brazilian territory. From the Atlantic Forest biome (latitude 22º), 14 specimens (seven males and seven female) were analyzed, and from the Pampa biome (latitude 29º), 20 specimens (eight males and twelve female). On average, in the right antimere the kidneys measured 49.9 ´ 25.2 ´ 24.4 mm, had a volume of 16.5 cm3, the renal artery measured 21.3 mm and the renal vein 19.4 mm. In the left antimere, the kidneys measured 49.3 ´ 24.4 ´ 22.8 mm, with a  volume of 14.6 cm3 and the artery and vein measured 21.0 mm and 28.4 mm, respectively. The right kidney was always cranial and predominantly positioned ventrally to vertebrae L1L3, while the left one was positioned ventrally to vertebrae L2L4. There was no difference in the comparison between sexes or antimeres. Most renal dimensions were significantly higher in the specimens from the Pampa biome, possibly due to the body size and type of diet.(AU)

Cerdocyon thousé o canídeo sul americano com maior distribuição geográfica. Ao sul do Equador, duas populações distintas são reconhecidas, submetidas a diferentes temperaturas médias e disponibilidade de alimento. Objetivou-se mensurar o comprimento, largura, espessura e volume dos rins, comprimento dos vasos renais e esclarecer a esqueletopia renal em duas populações isoladas de C. thous.Para tal, foram estudados rins e vasos renais de 34 cadáveres coletados mortos em rodovias brasileiras. Catorze espécimes (sete machos e sete fêmeas) são provenientes do bioma Mata Atlântica (latitude 22º) e 20 espécimes (oito machos e doze fêmeas) do bioma Pampa (latitude 29º). Na média, os rins direitos mediram 49,9x25,2x24,4 mm, tinham um volume de 16,5 cm3e a artéria renal direita mediu 21,3 mm e a veia renal direita 19,4 mm. Os rins esquerdos mediram 49,3x24,4x22,8 mm tinham volume de 14,6 cm3e as artérias e veias renais direitas mediram 21,0 mm e 28,4 mm, respectivamente. Os rins direitos foram sempre craniais e predominantemente posicionados ventralmente às vertebras L1 a L3, enquanto os rins esquerdos estavam posicionados majoritariamente ao nível de L2 a L4. Não houve diferença na comparação das medidas entre os sexos ou antímeros. Praticamente todasas medidas renais foram significativamente maiores nos espécimes do bioma Pampa, possivelmente devido(AU)

Comparative descriptive anatomy of the femoral artery in crab-eating fox, hoary fox and maned wolf / Anatomia descritiva comparativa da artéria femoral de cachorro-do-mato, raposa-do-campo e lobo-guará

The goal of this study was to describe the anatomy of the femoral artery in wild canids such as the crab-eating fox (Cerdocyon thous), the hoary fox (Lycalopex vetulus), and the maned wolf (Chrysocyon brachyurus). Two specimens from each canid group were used. Red dyed latex was injected in the arterial system of the animals, which were then fixed in 10% formaldehyde aqueous solution and dissected following the routine techniques in macroscopic anatomy. In the three canid groups, the arterial pattern was similar to that described for domestic canids, in which the deep femoral artery rises from the external iliac artery, still in the abdominal cavity, and sends its first branch, the lateral circumflex femoral artery. A few muscular branches, one or two caudal femoral arteries, and the terminal branches ­ the descending genicular artery, the saphenous artery, and the popliteal artery ­ are all originated from the femoral artery. The origin pattern of these vessels also shows similarities with those from domestic canids, sometimes forming trunks and occasionally rising individually. Thus, it can be concluded that the anatomical pattern of the femoral artery and its branches in wild canids shows similarities with that from domestic canids, but inherent variations in each species are also present.(AU)

O objetivo deste estudo foi descrever a anatomia da artéria femoral em canídeos selvagens, como o cachorro do mato (Cerdocyon thous), a raposa do campo (Lycalopex vetulus) e o lobo-guará (Chrysocyon brachyurus). Foram utilizados dois espécimes de cada grupo canídeo. Solução de látex vermelha foi injetada no sistema arterial dos animais, que foram então fixados em solução aquosa de formaldeído a 10% e dissecados seguindo as técnicas rotineiras da anatomia macroscópica. Nos três grupos canídeos, o padrão arterial foi semelhante ao descrito para canídeos domésticos, em que a artéria femoral profunda origina da artéria ilíaca externa, ainda na cavidade abdominal, e envia seu primeiro ramo, a artéria femoral circunflexa lateral. Alguns ramos musculares, uma ou duas artérias femorais caudais e os ramos terminais - a artéria genicular descendente, a artéria safena e a artéria poplítea - são originários da artéria femoral. O padrão de origem desses vasos também mostra semelhanças com as de canídeos domésticos, às vezes formando troncos e ocasionalmente originando individualmente. Assim, pode-se concluir que o padrão anatômico da artéria femoral e seus ramos em canídeos selvagens mostra semelhanças com a dos canídeos domésticos, mas variações inerentes em cada espécie também estão presentes.(AU)

Comparative descriptive anatomy of the femoral artery in crab-eating fox, hoary fox and maned wolf / Anatomia descritiva comparativa da artéria femoral de cachorro-do-mato, raposa-do-campo e lobo-guará

The goal of this study was to describe the anatomy of the femoral artery in wild canids such as the crab-eating fox (Cerdocyon thous), the hoary fox (Lycalopex vetulus), and the maned wolf (Chrysocyon brachyurus). Two specimens from each canid group were used. Red dyed latex was injected in the arterial system of the animals, which were then fixed in 10% formaldehyde aqueous solution and dissected following the routine techniques in macroscopic anatomy. In the three canid groups, the arterial pattern was similar to that described for domestic canids, in which the deep femoral artery rises from the external iliac artery, still in the abdominal cavity, and sends its first branch, the lateral circumflex femoral artery. A few muscular branches, one or two caudal femoral arteries, and the terminal branches ­ the descending genicular artery, the saphenous artery, and the popliteal artery ­ are all originated from the femoral artery. The origin pattern of these vessels also shows similarities with those from domestic canids, sometimes forming trunks and occasionally rising individually. Thus, it can be concluded that the anatomical pattern of the femoral artery and its branches in wild canids shows similarities with that from domestic canids, but inherent variations in each species are also present.(AU)

O objetivo deste estudo foi descrever a anatomia da artéria femoral em canídeos selvagens, como o cachorro do mato (Cerdocyon thous), a raposa do campo (Lycalopex vetulus) e o lobo-guará (Chrysocyon brachyurus). Foram utilizados dois espécimes de cada grupo canídeo. Solução de látex vermelha foi injetada no sistema arterial dos animais, que foram então fixados em solução aquosa de formaldeído a 10% e dissecados seguindo as técnicas rotineiras da anatomia macroscópica. Nos três grupos canídeos, o padrão arterial foi semelhante ao descrito para canídeos domésticos, em que a artéria femoral profunda origina da artéria ilíaca externa, ainda na cavidade abdominal, e envia seu primeiro ramo, a artéria femoral circunflexa lateral. Alguns ramos musculares, uma ou duas artérias femorais caudais e os ramos terminais - a artéria genicular descendente, a artéria safena e a artéria poplítea - são originários da artéria femoral. O padrão de origem desses vasos também mostra semelhanças com as de canídeos domésticos, às vezes formando troncos e ocasionalmente originando individualmente. Assim, pode-se concluir que o padrão anatômico da artéria femoral e seus ramos em canídeos selvagens mostra semelhanças com a dos canídeos domésticos, mas variações inerentes em cada espécie também estão presentes.(AU)

Morfologia da laringe em Cerdocyon thous (Linnaeus, 1766) / Laryngeal morphology in Cerdocyon thous (Linnaeus, 1766)

Cerdocyon thous (cachorro-do-mato) é o canídeo silvestre mais comum em território sul-americano. Objetivou-se com o presente trabalho descrever a morfologia macroscópica, microscópica e comparada da laringe nesta espécie. Para tal, as laringes de dezesseis espécimes (quatro machos e doze fêmeas) foram analisadas quanto à topografia, forma, morfometria, musculatura intrínseca e histologia. A laringe dispôs-se ventralmente ao áxis e foi constituída por uma cartilagem tireoide e outra cricoide (hialinas), uma epiglótica (elástica) e um par de ariteoides (mistas). Um par de cartilagens sesamoides foi identificado entre os processos corniculados e a lâmina da cricoide. A morfometria revelou que a tireoide é a maior cartilagem. Não houve sinais definitivos de dimorfismo sexual na laringe de C. thous. O epitélio predominante foi do tipo pavimentoso estratificado o qual sofreu transição para pseudoestratificado cilíndrico ciliado ao nível do terço caudal da tireoide e rostral da cricoide. A laringe de C. thous mostrou semelhança com a do cão doméstico, ainda que o formato das cartilagens tenha apresentado diferenças.

Cerdocyon thous (crab-eating fox) is the most common wild canid in South American territory. The aim of this paper was to describe macroscopic, microscopic and comparative morphology of the larynx of this species. Analyses of larynges of sixteen specimens (four males and twelve females) revealed the topography, conformation, morphometrics, intrinsic muscles and histological characteristics. The larynx was positioned ventral to the axis and comprised one cricoid and one thyroid cartilage (hyaline), one epiglottis (elastic) and a pair of arytenoids (mixed). A pair of sesamoid cartilages was identified between the corniculate processes and the cricoid lamina. Morphometry revealed that thyroid cartilage was the largest. There were no conclusive signs of sexual dimorphism in the larynx of this species. Squamous stratified epithelium predominated and underwent transition to ciliated pseudo-stratified at the level of caudal thyroid and rostral cricoid portions. By comparison, larynx of C. thous showed similarity with the domestic dog, although the shape of cartilages has shown some variation.

Morfologia da laringe em Cerdocyon thous (Linnaeus, 1766) / Laryngeal morphology in Cerdocyon thous (Linnaeus, 1766)

Cerdocyon thous (cachorro-do-mato) é o canídeo silvestre mais comum em território sul-americano. Objetivou-se com o presente trabalho descrever a morfologia macroscópica, microscópica e comparada da laringe nesta espécie. Para tal, as laringes de dezesseis espécimes (quatro machos e doze fêmeas) foram analisadas quanto à topografia, forma, morfometria, musculatura intrínseca e histologia. A laringe dispôs-se ventralmente ao áxis e foi constituída por uma cartilagem tireoide e outra cricoide (hialinas), uma epiglótica (elástica) e um par de ariteoides (mistas). Um par de cartilagens sesamoides foi identificado entre os processos corniculados e a lâmina da cricoide. A morfometria revelou que a tireoide é a maior cartilagem. Não houve sinais definitivos de dimorfismo sexual na laringe de C. thous. O epitélio predominante foi do tipo pavimentoso estratificado o qual sofreu transição para pseudoestratificado cilíndrico ciliado ao nível do terço caudal da tireoide e rostral da cricoide. A laringe de C. thous mostrou semelhança com a do cão doméstico, ainda que o formato das cartilagens tenha apresentado diferenças.(AU)

Cerdocyon thous (crab-eating fox) is the most common wild canid in South American territory. The aim of this paper was to describe macroscopic, microscopic and comparative morphology of the larynx of this species. Analyses of larynges of sixteen specimens (four males and twelve females) revealed the topography, conformation, morphometrics, intrinsic muscles and histological characteristics. The larynx was positioned ventral to the axis and comprised one cricoid and one thyroid cartilage (hyaline), one epiglottis (elastic) and a pair of arytenoids (mixed). A pair of sesamoid cartilages was identified between the corniculate processes and the cricoid lamina. Morphometry revealed that thyroid cartilage was the largest. There were no conclusive signs of sexual dimorphism in the larynx of this species. Squamous stratified epithelium predominated and underwent transition to ciliated pseudo-stratified at the level of caudal thyroid and rostral cricoid portions. By comparison, larynx of C. thous showed similarity with the domestic dog, although the shape of cartilages has shown some variation.(AU)