Detection of Assemblage A, Giardia duodenalis and Eimeria spp. in alpacas on two Maryland farms

Sixty-one fecal samples were collected from adult alpacas and crias (ages 10 weeks to 10 years) on two farms in central Maryland. The farms raised both suri (silky-haired) and huacaya (crimpy-haired) breeds. Females and crias were housed together on pasture, whereas older/breeding males were maintained on separate pastures. Samples were subjected to a density gradient centrifugation protocol to concentrate parasites and remove fecal debris and were examined by immuno-fluorescent and differential interference contrast microscopy. Oocysts of Eimeria spp. were noted in 14 fecal samples, 6 on MD-1 and 8 on MD-2. Based on oocyst morphometrics two species of Eimeria were present: E. punoensis (19.2 microm x 16.5 microm) and E. alpacae (23.7 microm x 19.5 microm). Five animals shed exclusively E. punoensis, seven shed exclusively E. alpacae, and two had mixed infections. The Eimeria infections were not associated with obvious clinical signs. To determine the presence of Cryptosporidium and Giardia species and genotypes, DNA was extracted from feces and subjected to PCR utilizing specific primers for the ssu-rRNA gene for both parasites. All PCR positive samples were further analyzed by DNA sequencing to identify the species or genotypes that were present. Assemblage A, G. duodenalis was detected in fecal samples from two alpacas on MD-1 and in one alpaca on MD-2. Assemblage E, G. duodenalis and Cryptosporidium spp. were not detected on either farm. Although the prevalence on these two farms was low, alpacas can harbor zoonotic G. duodenalis, and this should be borne in mind by persons interacting with the animals.     

Pseudorabies virus infection in mink: a host-specific pathogenesis

Pseudorabies virus (PRV) is an alphaherpesvirus that causes a neurological disease in many wild and domestic animals. The neuropathology elicited by PRV is quite consistent regardless of the host with the only exception of mink, in which it is characterized by a vasculopathy rather than by an encephalitis. In this study, we aimed to investigate the underlying pathogenic mechanism(s) of PRV infection in mink by using immunohistochemistry and laser capture microdissection (LCM) on material from naturally and experimentally infected animals. The inflammatory reaction induced by PRV was minimal or absent not only in the nervous system, where we identified a low number of macrophages and a few T lymphocytes, but also in the primary replication site, the oropharyngeal mucosa; however, the number of PRV-infected cells detected by immunohistochemistry was extremely high both in the peripheral mucosa and in the nervous tissue. On the other hand, the vascular pathology included parenchymal hemorrhages of various degrees and, in specific cortical areas of the brain, fibrinoid degeneration of the capillary walls. Detection of viral antigens by immunohistochemistry revealed infection of endothelial cells of capillaries situated both in the oropharyngeal mucosa and in the brain stem; the presence of PRV DNA in vessels was further demonstrated by PCR performed on LCM samples of brain capillaries. These results can be interpreted as supporting the idea that the different pathology of the disease in mink may be the consequence of an increased endotheliotropism of PRV in this species. Infection of the vessel wall may then lead to vascular pathology and impairment in endothelial cell function, resulting in a weak immune response to infection.     

Endotracheal stenting therapy in dogs with tracheal collapse

Tracheal collapse in dogs is a common respiratory disorder, typically presenting with a history of chronic cough, increasing respiratory difficulties, and episodes of dyspnoea. Medical treatment is the therapy of choice and surgical repair is considered when patients do not respond well. Minimally invasive endotracheal stenting is a promising new therapy under investigation, but there remain significant challenges to overcome potential complications. The purpose of this article is to provide a comparative overview of intra-luminal stenting of the trachea in human and veterinary medicine. The currently available stents and their potential clinical application to the veterinary patient will be discussed.     

Assessing Knowledge Production for Agrosilvopastoral Systems in South America

In recent decades agroforestry has undergone significant development in Latin America. South America generates the most scientific research on the topic in the region. We conducted a comprehensive review and analysis of knowledge production for South American agroforestry that includes livestock grazing, known as agrosilvopastoralism (AS), examining how different sociopolitical factors such as sources of funding, institutional priorities, and international cooperation can bias the direction and objectives of scientific research. We assessed the major attributes of scientific publications on the topic (25 articles per country; overall n = 210) and the potential factors underlying the processes of research and development in the region. The tree component was the most studied, while the livestock component received less attention. Studies were mainly focused on the production of goods and services (monetary or nonmonetary approaches), except in Brazil, where conservation was the major study objective. Stakeholders were involved in more than half of the studies (60%), and they were mostly ranchers and local producers. More than half (70%) of the studies offered recommendations based on their results, and such recommendations were mostly concerned with the management of agrosilvopastoral system components. In general, studies were led just as often by local as foreign first authors and coauthored by more than three people as part of interinstitutional working groups. Brazil, Argentina, Colombia, and Chile had more frequent cooperation among institutions and countries but mainly used their own funding. In contrast, Bolivia, Ecuador, and Peru had almost 100% of their studies supported by foreign countries (North America and Europe). Countries with greater internal funding of research generated more long-term studies. Besides this, two clear trends were detected: 1) conservation and social aspects were mainly supported by sources from external countries led by foreign principal investigators, and 2) production issues were supported from sources within countries and supported high levels of cooperation among institutions.     

Diagnostic validity and costs of pooled fecal samples and individual blood or fecal samples to determine the cow- and herd-status for Mycobacterium avium subsp. paratuberculosis

Two tests are used on a regular basis to detect Mycobacterium avium subsp. paratuberculosis (Map): ELISA and fecal culture. Fecal culture is considered more sensitive and specific but is costly and requires 3-4 months for results. Pooling of fecal samples of individual animals may reduce the high costs of fecal culture. The objective of the study was to investigate the diagnostic validity and costs for pooling of fecal samples in dairy farms relative to culture or an ELISA on individual samples to determine the cow- or herd-status for Map. Fifty fecal and blood samples per herd were collected in 12 Chilean dairy herds. The sensitivity of pooling was estimated given the pool-size, amount of shedding in the pool and the prevalence in the herd. The sensitivity of the pools relative to individual fecal culture was 46% (95% CI 29-63%) and 48% (28-68%) for pools of 5 and 10 cows, respectively. The sensitivity of the pools was lower in pools with low shedders (26 and 24% for pools of 5 and 10, respectively) than in pools with moderate or heavy shedders (>75% sensitivity). Pools of 10 cows are the better option to determine or monitor the herd status. A whole-herd ELISA is the least expensive way to determine the status of individual cows but has a lower Se and Sp than individual culture.     

Enhanced resistance to bacterial infection byErwinia carotovora subsp.atroseptica in transgenic potato plants expressing the attacin or the cecropin SB-37 genes

Blackleg and soft rot diseases, caused by the bacteriumErwinia carotovora, are among the diseases that cause important losses in culture and storage of potato. In this paper, we introduced bacterial resistance into potato, via genes encoding for proteins with antibacterial activity. For this purpose, potato clones were transformed either with the gene encoding the acidic attacin protein fromHyalophora cecropia, or with the gene encoding the cecropin analog peptide SB37. These clones were evaluated for soft rot and blackleg resistance, after inoculation with the bacterial strainErwinia carotovora subsp.atroseptica T7. Results reported in this paper indicate that a considerable percentage of the potato clones (15–22%) showed increased resistance to bacterial infection, revealed by reduced severity of blackleg or soft rot symptoms. Expression of the transgenes was demonstrated in some of the clones by Northern blot analysis. This is the first report indicating that expression of the gene encoding for an attacin protein and for the cecropin SB-37 peptide in transgenic potato confers increased resistance to bacterial infection.     

Acute Toxicity of Ammonia and Nitrite to White Shrimp Penaeus setiferus Postlarvae

Penaeus setiferus postlarvae were exposed to acute levels of ammonia, nitrite, and to a mixture of both by a short-term static method, The 24h, 48-h and 72-h LC50 values for un-ionized ammonia were 1.49, 1.21 and 1.12 mg/L NH₃-N (un-ionized ammonia as nitrogen), and 11.55, 9.38 and 8.69 mg/L ammonia-N (un-ionized plus ionized ammonia as nitrogen). The 24-h, 48-h and 72-h LC50 values for nitrite were 268.06, 248.84 and 167.33 mg/L nitrite-N (nitrite as nitrogen). Nitrite was much less toxic than ammonia. The joint effect of ammonia and nitrite on the postlarvae was synergistic at 48-h exposure and antagonistic after 72 h. Postlarvae of P. setiferus may be considered as organisms sensitive to ammonia and nitrite.     

Substrate selection and effect of different substrates on survival and growth of juveniles of the freshwater crayfish <Emphasis Type="Italic">Cherax quadricarinatus</Emphasis> (von Martens 1868) (Decapoda,Parastacidae)

Survival and growth of juveniles of the Australian freshwater crayfish Cherax quadricarinatus were evaluated in aquaria with four different substrates covering their bottom. The four substrates were plastic mesh, small stones, fine sand and bare glass without cover (control group). Two size classes of juveniles were evaluated: recently hatched or early (~0.02 g of body weight) and pre-fattening or advanced (~1 g of body weight). A group of twelve early or ten advanced juveniles were placed in each of the 24 aquaria. Each substrate had a total of six aquaria, three containing early juveniles and the other three containing advanced juveniles groups. Each aquarium had an excess small PVC pipes that served as shelters. After 12 weeks, early juveniles raised in small stones or fine sand had a statistically significant heavier body mass than the control group while the advanced juveniles did not show statistically significant differences in body weight. In a second experiment, ten early and ten advanced juveniles were individually placed in a circular glass aquarium divided into four sections each containing one of the same substrates used in the first experiment. Each individual was filmed for 24 h, and the time spent in each compartment was registered. Both sizes of juveniles showed a marked preference for the small stones. This experiment was repeated with animals used in the first experiment and observed the same selection pattern indicating that substrate selection is independent of body mass or previous acclimation to the different substrates.     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

Selecting and ranking areas for conservation of Aegla (Crustacea: Decapoda: Anomura) in southern South America integrating biogeography,phylogeny and assessments of extinction risk

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