SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis

Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.     

Neospora caninum infection as a cause of reproductive failure in a sheep flock

Neospora caninum has been detected only sporadically in cases of ovine abortion, and it has therefore traditionally been considered as an unimportant parasite in small ruminants. This study was carried out with the aim of identifying the pathogen causing serious reproductive problems on a commercial sheep farm. Sera from all rams and ewes tested negative for antibodies against Border disease virus, Schmallenberg virus and Coxiella burnetii, and infections by these agents were therefore ruled out. Nevertheless, seropositivity to N. caninum and/or Toxoplasma gondii was detected, although the seroprevalence was higher in the case of N. caninum. The percentage of lambings and the number of lambs per dam were significantly lower in ewes that were seropositive to N. caninum while no effect on these parameters was detected in ewes that were seropositive to T. gondii. There was also no evidence of infection by T. gondii in the foetal/lamb tissues analyzed by PCR and/or immunohistopathological techniques. On the contrary, the DNA of N. caninum was detected in 13 out of 14 foetuses/lambs descendant from dams seropositive to this parasite. Characteristic lesions caused by N. caninum and/or its antigen were also detected. Genotyping of the N. caninum DNA revealed only two closely related microsatellite multilocus genotypes. The results clearly demonstrate that infection by N. caninum was the cause of the low reproductive performance of this sheep flock.     

Overlooked impacts and challenges of the new European discard ban

Discards are the portion of animal and plant material in the catch that is dumped back at sea. The Common Fisheries Policy plan proposed by the European Commission for 2014–2020 presents a controversial goal: to enforce the landing of fishing discards as a measure to encourage their reduction. This historical and political decision will shape the future of the fishing exploitation in European Seas. Discards generated by European fleets are not negligible, and its reduction is an ecological, socioeconomical and moral imperative. However, it must be achieved through the reduction in discards at source and the promotion of selective and non‐destructive gears. We argue it is doubtful that this discard ban will result in an effective reduction of discards. The proposed measure may, in fact, negatively affect ecosystems at all levels of biological hierarchy by disregarding the Ecosystem‐Based Approach to Fisheries and the Precautionary Principle. It could negatively impact several species by increasing fishing mortality, also commercial species if discards are not accounted in the total allowable catch. Communities preying on discards will likely be affected. The role discards currently play in the energy turnover of current ecosystems will be modified and should be fully evaluated. The landing of discards will likely generate new markets of fishmeal due to the growing demands for marine living resources. The ban will require substantial public investment to deal with technical problems on board and to control and enforce. Therefore, this measure should be only implemented after rigorous scientific and technical studies have been developed.     

Rearing of pike-perch larvae using formulated diets – first success with starter feeds

The present study evaluated the performance of two commercial diets: AgloNorse (AN) and BioKyowa (BK), and two experimental, formulated diets based on casein (C) or casein plus casein hydrolysates (CH) in rearing of pike‐perch larvae (Sander lucioperca L.). All fish were 5 day old and control group was fed live Artemia nauplii. Fish were sampled periodically for histological comparison of morphological changes in the digestive tracts. Survival of fish fed Artemia nauplii, BK and AN was similar: 54.4%, 50.8% and 52.4%, respectively, while the fish fed formulated diets C and CH showed considerably lower survival: 28.4% and 21.6% respectively. After 5 weeks of rearing, the average body mass of fish ranged from 212±32 mg in Artemia fed group to 53.8±6.8 mg in C diet fed group. A considerable vacuolization of supranuclear zone in enterocytes of posterior intestine was observed in the larvae fed commercial diets. No anomalies in liver development were found. Hepatocytes of fish fed BK diet showed larger glycogen storage areas, compared with those occupied by lipids. The highest zymogen accumulation of pancreatic cytoplasm was observed in fish fed Artemia. In fish fed C and CH diets, anomalies in digestive system development were indicated by lower and less numerous intestinal folds, smaller hepatocytes, retarded development of gastric glands, and in CH group – also local fatty degeneration of liver.     

Haematologic and immunologic parameters of bullfrogs,Lithobates catesbeianus,fed probiotics

The effects of two probiotics (P₁–Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P₂–Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg^?1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin – and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P₁ fed at a dose of 10 g kg^?1 of diet and probiotic P₂ fed at 5 g kg^?1 of diet.     

Hemangiosarcoma of the third eyelid in a cat

A case of hemangiosarcoma of the third eyelid in a 15-year-old Domestic Short-haired cat is reported. A small red mass protruding from the anterior surface of the third eyelid in the left eye was present. Only mild bilateral conjunctivitis and nuclear sclerosis were revealed on initial ophthalmic examination. After anti-inflammatory treatment, surgery and cryotherapy were performed. On histopathologic examination the mass was located just beneath the conjunctival epithelium, suggesting that the conjunctiva was the tissue of origin. The tumor showed high cellularity and was composed of pleomorphic spindle and polygonal cells arranged in interlacing bundles and solid sheets. The presence of slit-like spaces between tumor cells containing erythrocytes demonstrated the vascular differentiation. Mitotic activity was moderate, but atypical mitoses were detected. The surgical area healed uneventfully and after 7 months there was no recurrence. To the authors' knowledge, this is the first report of conjunctival hemangiosarcoma of the third eyelid in a cat.     

Effects of long-term dietary supplementation of monensin or saccharomyces cerevisiae on blood acid-base and productive performance in growing feedlot steers

The aim of this study was to evaluate the effects of two dietary supplements (monensin and a live yeast culture) on acid-base balance in steers maintained in a commercial feedlot system, considering effects over the growing period (14 to 23 weeks of age). A 63-day feedlot study was performed using 42 double-muscled Belgian Blue steers. Steers were allotted randomly to one of the three study groups: (1) control group [no supplementation, C], (2) monensin supplementation [MON] at a concentration of 30 mg/kg (DM basis), and (3) live Saccharomyces cerevisiae strain supplementation [SACC] at a dose of 500 mg/kg (DM basis).Venous blood samples were collected for the measurement of acid-base parameters and L-lactate. Production parameters were also used as a complementary tool for understanding the internal changes associated with supplementation. Our results show that during the study period no statistical differences were observed between supplemented and control steers, although non-supplemented animals tended to gain more efficiently than those fed monensin or yeast. Nevertheless, taking into account blood parameters, these control animals showed a greater risk of acid overload due to a more marked decline in blood buffer levels over time in comparison with supplemented steers although no differences were observed between monensin or yeast supplemented animals. Additionally, significant effect of supplementation was observed in packed cell volume (PCV) values.     

Effect of nutritional supplementation upon pregnancy rates of goats under semiarid rangelands and exposed to the male effect

The aim of this study was to evaluate possible effects of improved protein and energy diet upon reproductive outcomes of adult goats under marginal rangeland grazing conditions and exposed to the male effect during the anoestrous-dry season. Adult goats (Nubian × Criollo, nonpregnant, nonlactating, n?=?160, 22° 50' N, 100° 59' W, 1,653?m) were randomly divided in two experimental groups: Control (CG; n?=?80) and Supplemented (SG; n?=?80). Both CG and SG goats were kept together during the day for ranging a semiarid rangeland while separated accordingly at night. SG goats were individually supplemented [400?g?kg(-1)?day(-1)?; 14% CP and 2.92 Mcal DE kg(-1)] during a 30-day period. Thereafter, both the CG [35.5?±?0.94?kg LW, 1.95?±?0.06 U, body condition score (BCS)] and the SG [39.5?±?0.94?kg LW, 2.29?±?0.06 BCS] were exposed to adult males (Nubian, n?=?4, two per group) of proven libido and fertility and kept together at night (1700-0900) during a 42-day experimental breeding period. At the beginning of the breeding period, both LW (P??0.05), respectively. Results suggest that nutritional supplementation and the male effect were able to successfully invoke neurophysiological pathways to increase ovarian activity and to promote a uterine milieu prone to the establishment of pregnancy during the anoestrus season. However, the increased abortion rates observed in the study occurred irrespective of the nutritional regime offered during the premating stage. Therefore, there is a need to align the last third of gestation to the onset of the grazing season in order to increase reproductive and economic efficiency in those rangeland-based marginal goat production systems.     

Jaagsiekte sheep retrovirus found in milk macrophages but not in milk lymphocytes or mammary gland epithelia of naturally infected sheep

Jaagsiekte sheep retrovirus (JSRV) causes ovine pulmonary adenocarcinoma. JSRV can be transmitted via infected colostrum or milk, which contain somatic cells (SCs) harboring JSRV provirus. Nevertheless, the cell types involved in this form of transmission and the involvement of the mammary gland remain unknown. We separated adherent cells (macrophages and monocytes) by plastic adherence, and lymphocytes (CD4+ and CD8+ T cells, and B cells) by flow cytometry, from SCs in milk samples from 12 naturally infected, PCR blood test JSRV–positive, subclinical ewes. These cell populations were tested by PCR to detect JSRV provirus. The ewes were euthanized, and mammary gland samples were analyzed immunohistochemically to detect JSRV surface protein. We did not detect JSRV provirus in any milk lymphocyte population, but milk adherent cells were positive in 3 of 12 sheep, suggesting a potential major role of this population in the lactogenic transmission of JSRV. Immunohistochemistry did not reveal positive results in mammary epithelial cells, pointing to a lack of participation of the mammary gland in the biological cycle of JSRV and reducing the probability of excretion of free viral particles in colostrum or milk.