Hypoosmotic Swelling Test in Alpaca (Vicugna pacos) Epididymal Spermatozoa

The present study intended to develop the hypoosmotic swelling (HOS) test in alpaca for its use in epididymal spermatozoa, to evaluate the membrane functional integrity and determine an appropriate hypoosmotic solution and whether the incubation time of 15 or 60 min is sufficient for the execution of the test. Hypoosmotic solutions (HS) with the following concentrations were used: 50, 100, 150, 200 and 275 mOsm/kg of sodium citrate tribasic dihydrate and d ‐fructose. Ten microlitres of epididymal sperm sample was mixed in 150 μL of the respective HS and incubated for 15 or 60 min at 38°C. From the proportion of reacted (swollen) spermatozoa, the 150 mOsm/kg HS was the most sensitive (p < 0.05). The exposure times (15 and 60 min) did not have significant differences (p > 0.05) in the proportion of both strong‐ and total‐coiled sperm tails. In conclusion, 150 mOsm/kg HS and 15 min exposure time are optimal to evaluate the plasma membrane functional integrity through the HOS test in alpaca epididymal spermatozoa.     

Biochemical and Protein Profile of Alpaca (Vicugna pacos) Uterine Horn Fluid During Early Pregnancy

South American camelids show high embryo loss rate, during the first 60 days of pregnancy. One of the factors which may be related to this situation is that over 98% of the embryos implant in the left uterine horn (LUH) even though both ovaries contribute similarly to ovulation. There is scarce information about the uterine environment of female camelids at any physiological state that could explain the capability of the LUH to attract the embryo and maintain pregnancy. We describe, for the first time, the biochemical and protein profile of uterine fluid (UF), addressing the right and LUH environment in non‐pregnant and pregnant alpacas. Different substrates, electrolytes and metabolites were assayed in both uterine horn fluids. Small changes were observed in glucose and total protein levels, which were more noticeable during pregnancy. In addition, 10 specific proteins were found in the left horn fluid in 5‐week‐pregnant alpacas, and two protein bands were identified in non‐pregnant alpaca right horn fluid. These results would provide basic information for identification of possible markers for pregnancy diagnosis, reproductive diseases and hormone‐treated animals evaluation and hence contributing to improve the pregnancy rate.     

Presence and Distribution of Urocortin and its Receptors in the Epididymis of Alpaca (Vicugna pacos)

Urocortin 1 (UCN) is a 40‐amino acid peptide belonging to the corticotrophin‐releasing hormone (CRH) family. The biological effects of this peptide are modulated by binding two G‐coupled receptors named CRH receptor 1 (CRHR1) and CRH receptor 2 (CRHR2). CRHR2 has high affinity for UCN. The aim of the present study was to investigate the presence and distribution of UCN, CRHR1 and CRHR2 in the epididymis of the South America camelid Alpaca (Vicugna pacos) by Western blotting analysis and immunohistochemistry. Tissue extracts of the organ reacted with the anti‐UCN, anti‐CRHR1 and anti‐CRHR2 antibodies, recognizing in all the cases a single specific protein band. UCN‐ and CRHR2‐immunoreactivities (IRs) were found in the cytoplasm of the principal cells (PCs) of the caput epididymis. A prevalent supranuclear localization of granular‐shaped positive material was observed. CRHR1‐IR was observed in the fibromuscular stromal cells encircling the tubules and in the smooth musculature of the blood vessels throughout the three epididymal segments. In addition, in the cauda, CRHR1‐IR was observed in some apical epithelial cells (ACs) which were morphologically similar to apical mitochondria‐rich cells (AMRCs). These results suggest that UCN, CRHR1 and CRHR2 are expressed in the alpaca epididymis and that CRH‐related peptides might play multiple roles in maturation and storage of spermatozoa.     

Expression of Orexin A and its Receptor 1 in the Epididymis of the South American Camelid Alpaca (Vicugna pacos)

Orexins A (ox A) and B are two peptides originally discovered in neurons of rat hypothalamus, and later found in different cellular types of the gastrointestinal and genital tracts. They arise from the proteolytic cleavage of a common precursor molecule, prepro‐orexin, and bind to two receptors, namely receptor 1 (ox1r) and receptor 2 for orexins, that show different binding affinity. The central role of the two peptides has been extensively studied, whereas their activity in the periphery is still poorly known. Here, we investigated the presence of ox A and ox1r in the epididymis of a South American camelid species, the alpaca, by immunohistochemistry, and we also assessed the expression of prepro‐orexin and ox1r in tissue extracts by Western blotting analysis. Ox A‐ and ox1r‐immunoreactivity was found in the cytoplasm of principal cells of the caput epididymis. A prevalent supranuclear localization of granular‐shaped positive material was observed. No positivity was present in the other cytotypes of epididymis. The expression of two peptides with molecular weight corresponding to those of prepro‐orexin and ox1r, respectively, was detected in the tissue extracts from the organ.     

Diaphragmatic hernia in a two-year-old alpaca (Vicugna pacos)

A 2-yr-old male intact alpaca (Vicugna pacos) was admitted for a 4-day history of anorexia and colic. Five months prior, the alpaca had undergone surgical removal of a duodenal trichophytobezoar and had recovered uneventfully. The alpaca died under anesthesia, and diaphragmatic herniation of the third gastric compartment (C3) was diagnosed at necropsy. A defect was identified in the left dorsal hemidiaphragm accompanied by herniation of 80% of C3 and the aboral portion of the second gastric compartment into the pericardial sac. The smooth margins and dorsal location of the diaphragmatic defect suggested a congenital origin. Diaphragmatic herniation is uncommon in camelids, and only one other case has been reported. Due to the dorsal location of the diaphragmatic defect in this animal, positioning during the previous surgery may have initiated a partial entrapment of gastric compartments, leading to a more complete incarceration between when the animal was discharged and presented again.     

Multiple follicular cysts in four alpacas (Vicugna pacos)

Follicular cysts are non-neoplastic skin lesions lined by follicular epithelium. The pathogenesis of these lesions is unclear. Multiple follicular cysts occur sporadically in dogs, horses and humans and are common in sheep. Here we report multiple follicular cysts in four aged alpacas (Vicugna pacos). Based on their histological features, they are most consistent with hybrid cysts. This is the first report of multiple follicular cysts in alpacas.     

Pulmonary alveolar microlithiasis in an alpaca (Vicugna pacos)

Pulmonary alveolar microlithiasis (PAM) is a rare, familial disease of unknown aetiology characterised by intra‐alveolar formation and accumulation of microliths. Multiple formalin‐fixed tissues were submitted from a 5‐month‐old female alpaca that died suddenly without significant clinical signs. No gross abnormalities were observed on postmortem examination. Histological findings included PAM and severe centrilobular hepatic necrosis. Although the hepatic lesion was the likely cause of death, PAM was an incidental finding that has not been reported previously in alpacas. An overview of PAM, including pathogenesis and histopathological characteristics, are discussed in relation to the concurrent hepatic disease in the present case.     

Mapping of Neurotensin in the Alpaca (Lama pacos) Brainstem

We studied the distribution of cell bodies and fibres containing neurotensin (NT) in the brainstem of the alpaca using an indirect immunoperoxidase technique. Immunoreactive fibres were widely distributed throughout the brainstem, whereas the distribution of cell bodies was less widespread. Immunoreactive perikarya were only found in the mesencephalic and bulbar reticular formation, periaqueductal grey, nucleus of the solitary tract, laminar spinal trigeminal nucleus and in the inferior colliculus. A high density of fibres containing NT was found in the dorsal nucleus of the raphe, marginal nucleus of the brachium conjunctivum, locus coeruleus, inferior colliculus, inter‐peduncular nucleus, substantia nigra, periaqueductal grey, reticular formation of the mesencephalon, pons and medulla oblongata, nucleus of the solitary tract, laminar spinal trigeminal nucleus, hypoglossal nucleus, inferior central nucleus and in the tegmental reticular nucleus. The widespread distribution indicates that NT might be involved in multiple physiological actions in the alpaca brainstem; this must be investigated in the future as alpacas lives from 0 m above sea level to altitudes of up 5000 m and hence the involvement of this neuropeptide in special and unique regulatory physiological mechanisms could be suggested.     

Histological characterization of umbilical cord in alpaca (Vicugna pacos)

The histomorphometric features of umbilical cord constituents in seven foetuses of alpaca (Vicugna pacos) from Cerro de Pasco, Department, Peru, were determined. Sections of 2–5 cm of umbilical cord were fixed in 10% neutral buffered formalin and processed for light microscopy. Standard histological slides stained with haematoxylin and eosin, Masson's trichrome and Van Gieson's trichrome were obtained. Histologically, common features of umbilical artery and vein were observed as well as mucous connective tissue, some cell features that compound this tissue constituted by cells presented features of myofibroblasts. Among most important findings that were observed, the lumen of umbilical vein was obliterated into star‐shaped form with the thinner umbilical artery wall; the smooth muscles and fibroblast were comparatively more in number in umbilical artery than that of umbilical vein, and the tunica media was larger in dimension than the tunica adventitia in umbilical vein. Conclusively, this histological study features an observation of the umbilical cord of alpaca foetuses and shows the similarity between them and those of other mammal species, including dromedaries and South American camelids.     

Anthelmintic resistance in a herd of alpacas (Vicugna pacos)

A herd of alpacas was examined because of a history of severe endoparasitism, anemia, hypoproteinemia, and weight loss. Resistance of gastrointestinal nematodes to albendazole, fenbendazole, and doramectin was documented. This report suggests that anthelmintic resistance may be an emerging problem in South American camelids in North America.     

Bilateral iliac arterial thrombosis in an alpaca (Vicugna pacos) cria

A 3-month-old male alpaca was presented for acute onset of non-weight-bearing left hind limb lameness. Antemortem diagnostics revealed a mass to the right of the urinary bladder and no blood flow in the left femoral artery. Necropsy revealed bilateral iliac arterial thrombi and an abscess near the urinary bladder.