Luis Eusebio Fidalgo

Seropathotypes, Phylogroups, Stx Subtypes, and Intimin Types of Wildlife-Carried, Shiga Toxin-Producing Escherichia coli Strains with the Same Characteristics as Human-Pathogenic Isolates

ABSTRACT The objectives of this study were to investigate the presence of Shiga toxin-producing Escherichia coli (STEC) strains in wildlife that have spread in Europe, living near human settlements; to analyze their epidemiological role in maintenance and transmission to domestic livestock; and to assess the potential health risk of wildlife-carried strains. STEC strains were recovered from 53% of roe deer, 8.4% of wild boars, and 1.9% of foxes sampled in the northwest of Spain (Galicia). Of the 40 serotypes identified, 21 were classified as seropathotypes associated with human disease, accounting for 81.5% of the wildlife-carried STEC strains, including the enterohemorrhagic serotypes O157:H7-D-eae-γ1, O26:[H11]-B1-eae-β1, O121:H19-B1-eae-ε1, and O145:[H28]-D-eae-γ1. None of the wildlife-carried strains belonged to the highly pathogenic serotype O104:H4-B1 from the recent Germany outbreak. Forty percent of wildlife-carried STEC strains shared serotypes, phylogroups, intimin types, and Stx profiles with isolates from human patients from the same geographic area. Furthermore, wildlife-carried strains belonging to serotypes O5:HNM-A, O26:[H11]-B1, O76:H19-B1, O145:[H28]-D, O146:H21-B1, and O157:H7-D showed pulsed-field gel electrophoresis (PFGE) profiles with >85% similarity to human-pathogenic STEC strains. We also found a high level of similarity among STEC strains of serotypes O5:HNM-A, O26:[H11]-B1, and O145:HNM-D of bovine (feces and beef) and wildlife origins. Interestingly, O146:H21-B1, the second most frequently detected serotype in this study, is commonly associated with human diarrhea and isolated from beef and vegetables sold in Galicia. Importantly, at least 3 STEC isolates from foxes (O5:HNM-A-eae-β1, O98:[H21]-B1-eae-ζ1, and O146:[H21]-B1) showed characteristics similar to those of human STEC strains. In conclusion, roe deer, wild boar, and fox in Galicia are confirmed to be carriers of STEC strains potentially pathogenic for humans and seem to play an important role in the maintenance of STEC.

Bioaccumulation of cadmium, lead and zinc in liver and kidney of red fox (Vulpes vulpes) from NW Spain: influence of gender and age

Cd, Pb, and Zn were quantified in liver and kidney of red foxes (Vulpes vulpes) which were hunted during the 2003–2011 hunting seasons in Galicia (NW Spain). The effects of age and gender were evaluated to determine whether these variables should be included in future biomonitoring studies. The concentrations of hepatic and renal Cd (average 0.6 and 1.3 µg/g) and Pb (0.8 and 0.06 µg/g, respectively) were similar to background levels, with no known toxicological relevance. Similarly, the average levels of Zn in liver and kidney (77 and 17 µg/g) were in the range of physiological levels for canids. Although no significant gender-dependent variations were observed, the effect of aging was evident: the levels of hepatic Pb and both hepatic and renal Cd were higher in adults than in juveniles. Age should be included as a parameter during future biomonitoring programs focusing on trace metal bioaccumulation in red foxes.

Calbindin D28k Expression and the Absence of Apoptosis in the Cerebellum of Solanum Bonariense L-Intoxicated Bovines

Solanum bonariense intoxication is characterized by cerebellar neuronal vacuolation, degeneration, and necrosis. Cerebellar Purkinje cells seem especially susceptible, but more research is needed to determine the pathogenesis of neuronal necrosis and the mechanism of Purkinje cell susceptibility. Calbindin D28k (CbD28k) is highly expressed in Purkinje cells and has been used as a marker for normal and degenerative Purkinje cells. The goal of this study was to describe S bonariense–induced disease by ascertaining Purkinje cell–specific degenerative changes using CbD28k expression and to correlate this with apoptosis in Purkinje cells, as determined using TUNEL (transferase-mediated dUTP-biotin nick end-labeling) and ultrastructural changes. In all cases, an increase in both dose and duration of S bonariense intoxication resulted in a decrease in the number of Purkinje cells. CbD28k immunohistochemistry was an excellent marker for Purkinje cells because immunoreactivity did not change in normal or degenerative tissues. This finding suggests that excessive calcium excitatory stimulation does not induce rapid neuronal degeneration and death. As found in previous studies, TUNEL tests and electron microscopy suggest that Purkinje cell degeneration and death are not occurring via an apoptotic process. These findings suggest that S bonariense poisoning induces progressive Purkinje cell death that is not mediated by excitotoxicity or apoptotic activation.

Quantification of pericaudal adipocyte diameter in dairy cattle during peripartum: a complementary method to study energetic status using conventional histology

The energetic status of high-yielding Holstein-Friesian dairy cattle was studied during peripartum under field conditions using body condition score (BCS), glycemia, seric ß-hydroxybutyrate and adipose tissue cellularity. This last method was tested as a complementary tool for energetic status assessment. Biopsies of pericaudal subcutaneous adipose tissue were obtained from 25 multiparous animals at 28 days before and 21 days after parturition. Samples were routinely processed for histological examination and stained with haematoxylin and eosin (H&E). The mean diameter of adipocytes (MDA) was measured with the aid of a digital image processor. During the same period, blood samples were collected weekly for metabolite determinations. The MDA at 28 days pre-partum and 21 days post-partum were 72.1 vs. 66.2 μm respectively (p = 0.055), and the corresponding BCS at these moments was 3.32 vs. 3.19 (p = 0.068). At -28 days pre-partum, the BCS was positively correlated with MDA (Pearsons r = 0.521, p = 0.016) and with glycemia (Pearsons r = 0.404, p = 0.056). Correlations between BCS and MDA, and between BCS and glycemia, with ß-hydroxybutyrate although not significant, suggest that routine histological preparations of biopsies from subcutaneous adipose tissue could be included as an easy and valuable tool for research purposes to evaluate metabolic adaptation of dairy cows to peripartum, as well as the incidence of metabolic disorders and productive performance.

Babesia microti-like piroplasm (syn. Babesia vulpes) infection in red foxes (Vulpes vulpes) in NW Spain (Galicia) and its relationship with Ixodes hexagonus

Piroplasmosis is caused by several species of protozoa such as the Babesia microti-like piroplasm (Bml), an emerging blood protozoan also known as Theileria annae or Babesia vulpes. Infection by Bml was first reported in dogs in Spain where it is endemic today. Recently, a high prevalence of Bml has been increasingly detected in red foxes (Vulpes vulpes) in European countries. The objective of this study was to determine infection levels of this parasite in foxes from Galicia, NW Spain, and ticks species infestation in these carnivores, where they are so far unknown. Samples of blood, spleen and ticks (if present) were taken from 237 hunted red foxes in the Galicia region. Blood smears were prepared for direct parasite observation, and spleen and tick samples were examined by nested PCR. Prevalences of Bml infection in Galician red foxes were estimated at 72% (171/237) by PCR and 38.23% (26/68) by direct observation. Among 837 ticks collected, the main tick identified was Ixodes hexagonus (present in 82.4% of the foxes) followed by Ixodes ricinus (12.3%), Dermacentor reticulatus (12.3%) and Rhipicephalus sanguineus sensu lato (3.5%). From 34 foxes testing positive for Bml, 616 ticks were collected: positive Bml PCR results were obtained in 55.6% (227/408) of ticks collected from 9 foxes, while the 208 ticks from the remaining 25 infected foxes returned negative PCR results. Given that canine piroplasmosis is endemic in this area, our observations point to the red fox as the main reservoir for Bml infection and the high proportion of I. hexagonus among ticks collected from red foxes suggests its likely role as vectors of B. microti-like piroplasm in this region. Further studies are needed for a better understanding of the link between the wild and domestic life cycles of this piroplasm.

The American mink (Neovison vison) is a competent host for native European parasites

The American mink (Neovison vison) is a mustelid native to North America that was introduced in Europe and the former USSR for fur farming. Throughout the last century, accidental or deliberate escapes of mink from farms caused the establishment of stable feral populations. In fact, the American mink is considered an invasive alien species in 28 European countries. The present study evaluates the gastrointestinal and cardiopulmonary helminth fauna of the American mink in Galicia (NW Spain) to understand its role as a potential reservoir for parasites affecting other autochthonous mustelids. In the period 2008-2014, fifty American mink (35 males and 15 females) of different ages (22 immature and 28 adults) from the provinces of Lugo, Ourense and Pontevedra were captured and sacrificed. Eight parasite species were found (6 nematodes and 2 trematodes) with the following prevalences: Molineus patens (68%), Aonchotheca putorii (54%), Crenosoma melesi (10%), Aonchotheca annulosa (8%), Angiostrongylus daskalovi (6%), Aelurostrongylus spp. (2%), Troglotrema acutum (2%) and an unidentified trematode (2%). Eighty-two per cent of the mink harboured helminths, including 15 animals (30%) infected by only one parasite species, 19 (38%) by two species, 5 (10%) by three species and 2 mink (4%) by four species. All helminth species identified are native to European mustelids. Statistical models were used to evaluate if animal characteristics (age, sex and weight), date and capture area influenced the prevalence, intensity or parasite richness. Statistical differences were detected only in models for intensity of M. patens, A. putorii and C. melesi. This is the first report of Angiostrongylus daskalovi, a cardiopulmonary nematode, and A. annulosa, a gastrointestinal nematode specific of rodents, in American mink. Moreover, although the fluke T. acutum has already been cited in American mink, to our knowledge, the present study represents the first report of this trematode in the lung.

Utilización de las sales aniónicas en la prevención de la paresia puerperal hipocalcémica

SUMMARY Milk fever is one of the most common metabolic diseases of dairy cattle. Of the various methods used in attempting to control the disease, most progress has been made in terms of dietary management. Many authors have shown that adding anionic salts to a ration has dramatic effects on the incidence of milk fever. Anionic salts are added to the diet of dry cows 2-3 weeks before calving in order to achieve metabolic acidosis. The exact mechanism of how dietary anions work is still unresolved. Previous reports suggested that metabolic acidosis induced by anionic salts increases tissue responsiveness to parathyroid hormone, ameliorates the absorption of calcium in the intestine and enhances calcium resorption from bone. In this work, we review the concept of dietary cation-anion difference (DCAD) with special emphasis on the physiologic effects this can have on acid-base status, bone resorption, renal production of 1,25 (OH) 2 D 3 and resorption of calcium, and intestinal absorption of calcium in cows.