Genetic analysis of human and swine influenza A viruses isolated in Northern Italy during 2010–2015

Influenza A virus (IAV) infection in swine plays an important role in the ecology of influenza viruses. The emergence of new IAVs comes through different mechanisms, with the genetic reassortment of genes between influenza viruses, also originating from different species, being common. We performed a genetic analysis on 179 IAV isolates from humans (n. 75) and pigs (n. 104) collected in Northern Italy between 2010 and 2015, to monitor the genetic exchange between human and swine IAVs. No cases of human infection with swine strains were noticed, but direct infections of swine with H1N1pdm09 strains were detected. Moreover, we pointed out a continuous circulation of H1N1pdm09 strains in swine populations evidenced by the introduction of internal genes of this subtype. These events contribute to generating new viral variants—possibly endowed with pandemic potential—and emphasize the importance of continuous surveillance at both animal and human level.     

Effects of energy level and protein source on nitrogen kinetics in steers fed wheat straw-based diets.

A 4 x 4 Latin square metabolism trial with a 2 x 2 factorial arrangement of treatments was conducted to determine N kinetics in steers. Steers were fed either untreated (UNT-WS) or alkaline hydrogen peroxide-treated wheat straw (AHP-WS) based diets supplemented with soybean meal (SBM) or blood meal (BM). Single doses of (15NH4)2SO4 were infused into ruminal pools to determine N kinetics. Ruminal NH3N concentrations (main effects) were 3.81, 1.65, 3.18, and 2.28 mg/dL in steers when fed diets that contained UNT-WS, AHP-WS, SBM, and BM, respectively. Ruminal N pool size was greater (P < .05) for UNT-WS than for AHP-WS diets and also was greater (P < .10) for SBM than for BM diets. Nitrogen flux rate into the rumen was not affected (P > .10) by diet. However, production rate of N from the ruminal pool was greater (P < .05) for UNT-WS than for AHP-WS diets and greater (P < .10) for SBM than for BM diets. Nitrogen recycled into the rumen was 33% greater (P < .05) for AHP-WS than for UNT-WS diets and 26% greater (P < .05) for BM than for SBM diets. Nitrogen recycling (percentage of N intake) was 33, 56, 36, and 49% for UNT-WS, AHP-WS, SBM, and BM diets, respectively. The blood urea N (BUN) concentrations were 10.23, 4.58, 7.15, and 7.65 mg/dL for UNT-WS, AHP-WS, SBM, and BM diets, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Adjuvant enhancement of humoral immune response to chemically inactivated bovine viral diarrhea virus.

Potentiation of the antibody response to inactivated bovine viral diarrhea virus by immunological adjuvants was studied in guinea pigs and cattle. The inactivated bovine viral diarrhea virus alone was demonstrated to be a weak immunogen. Addition of either 2 mg per mL diethylaminoethyl-dextran or 5% alhydrogel to inactivated bovine viral diarrhea virus did not or only slightly stimulated the antibody response; the combined adjuvants induced a significantly higher titer. A higher concentration (20 mg per mL) of diethylaminoethyl-dextran, on the contrary, suppressed the immune potentiation by the dual adjuvants. Combination of Bordetella bronchiseptica and alhydrogel adjuvants stimulated a high titer of antibody. The titer was further elevated upon revaccination and was significantly higher than that induced by alhydrogel alone. In cattle, alhydrogel enhanced the immune response and the additional inclusion of diethylaminoethyl-dextran did not cause a significant potentiation of the immunity. However, the antibody decay rate was significantly slower when stimulated by the combined adjuvants.     

Glasser's disease and prevalence of subclinical infection with Haemophilus parasuis in swine in southern Ontario

Glasser's disease accounted for less than 1% of total swine mortalities in an 11 year retrospective postmortem survey of swine submissions at three provincial government diagnostic laboratories in southern Ontario. However, Glasser's disease was suspected in 17 of 83 boar mortalities at the Record of Production Boar Test Station between 1983 and 1985 and was much more common in specific-pathogen-free (SPF) boars than in conventional boars. The prevalence of the causative organism, Haemophilus parasuis, was determined for 19 SPF herds in Ontario classified as "Excellent" under the Ontario Swine Herd Health Policy. Nasal swabs from two-month-old pigs were cultured on chocolate agar containing 1.5 mug/mL lincomycin, 5 mug/mL bacitracin, and 0.1 mug/mL crystal violet. Three herds were negative for H. parasuis infection; 16 herds contained clinically healthy carrier pigs.     

Effect of liming on extractable Zn, Cu, Fe and Mn in selected Scottish soils

Ten naturally acidic Scottish soils have been limed with CaCO₃ at 0,0.25, 0.50, 0.75, 1.00 and 1.25 times the recommended lime requirement, and then subjected to three cycles of alternate wetting to field capacity at 20°C for 1 month and drying for 1 month. The Zn, Cu, Fe and Mn extracted by DTPA solution, both at pH 7.3 and also after adjustment to match the final soil pH, were measured by flame AAS. The same elements were determined in ammonium acetate extracts, for extractants buffered at pH 4.8, pH 7.0 or at the final soil pH. The effect of extractant pH upon the amounts of trace elements removed is discussed for the two extractants, with particular reference to Zn. The influence of soil pH upon the amounts of trace elements extracted is also considered. There appears to be a distinct relationship between rate of decrease of DTPA-extractable Fe with increasing soil pH and the effect of pH upon DTPA-extractable Zn, high rates of decrease of extractable Fe being associated with a more continuous decline in extractable Zn with increasing soil pH. Possible reasons for this relationship are discussed, in the light of an experiment conducted to examine the effect of Fe and A1 on loss of Zn from solution at high pH. DTPA-extractable Cu varied little with increasing soil pH.     

Effects of graded levels of potato by-products in barley- and corn-based beef feedlot diets: II. Palatability

The objective of this study was to evaluate the effects of barley- or corn-based diets containing 0, 10, or 20% potato by-product (DM basis) on Warner-Bratzler shear force and palatability of beef. One hundred forty-four crossbred beef steers (333+/-.44 kg) were allotted within weight block (3) to a randomized complete block design with a 2 x 3 factorial arrangement of dietary treatments. Main effects were grain (barley or corn) and level of potato by-product (0, 10, or 20% of diet DM). There were a total of 18 pens with eight steers per pen and three pens per treatment. Steers were fed diets containing 83% concentrate (grain plus potato by-product), 10% supplement, and 7% alfalfa (DM basis) for an average of 130 d. Longissimus muscle cuts were used for Warner-Bratzler shear force determination (four steers per pen) and evaluation (two steers per pen) by a 10-member trained laboratory panel, a professional flavor/texture profile panel, and by consumer panels. Diet did not affect (P > .10) Warner-Bratzler shear force or trained laboratory panel tenderness, juiciness, and flavor intensity scores. Flavor/texture profile panel scores indicated feeding a corn-based diet as opposed to barley-based diet produced a more appropriate well-balanced and well-blended beef flavor and texture. However, the magnitudes of the differences were relatively small, and flavor and texture amplitude ratings for both barley- and corn-fed beef were well above average. Beef from steers fed 10 or 20% potato by-product had lower (P < .05) incidences of inappropriate aromatics and aftertastes, which may have a slightly beneficial effect on beef flavor, but flavor amplitude was not affected (P > .05) by level of potato. Moreover, consumer panel overall acceptability scores were not affected by diet. Thus, feedlot diets containing corn or barley with or without potato by-product should result in palatable beef products.     

Canine bronchomalacia: a clinicopathological study of 18 cases diagnosed by endoscopy

Canine bronchomalacia (BM) is characterized by weakness leading to collapse of the bronchial wall. A prospective study of 18 affected dogs (age range: 1-15 years) was undertaken to characterize the clinicopathological and histological features of BM. Poodles and Yorkshire terriers were commonly affected. Half of the dogs were overweight or obese. The clinical presentation was a mild, wheezing, chronic cough and pulmonary crackles were heard in 28% of the dogs. Compatible radiographic changes were present in 61% of the dogs. Using bronchoscopy, both lungs were affected in half of the animals, whereas in the others the disease appeared to affect predominantly the left lung. Analysis of bronchoalveolar lavage fluid and biopsies of bronchial mucosa revealed pure or mixed neutrophilic inflammation. Underlying infectious bronchitis was considered possible in 56% of the dogs. It was concluded that canine BM may present as an isolated clinical entity associated with infection and/or inflammation.     

FC‐14 Clonality studies of feline cutaneous lymphocytosis

A previous study described cutaneous lymphocytosis (CL) in 23 cats. The process resembles cutaneous pseudolymphoma in humans, a heterogeneous group of benign reactive proliferations of well‐differentiated lymphocytes in the skin of humans. Morphological and immunophenotypic characteristics do not offer reliable criteria to accurately predict the clinical outcome of feline CL or pseudolymphoma in humans. Presence of clonal cell populations is more consistent with a neoplastic process. In a previous study, feline CL lesions (20 cats) were evaluated for clonality using PCR, and only two cats had monoclonal T‐cell populations. Because false‐negative results may occur, the purpose of this study was to repeat the PCR using a revised primer set based on analysis of additional feline T‐cell receptor γ (TCRγ) sequences. DNA was isolated from 29 skin lesions and six internal organs of 20 cats. DNA integrity was assessed by glyceraldehyde‐3‐phosphate dehydrogenase PCR. Polymerase chain reaction clonality was performed using the revised primer set specific for feline TCRγ, and duplicate samples were evaluated. The PCR products were assessed by heteroduplex analysis. Clonal rearrangement of TCRγ was detected in 14 cats (24 of 35 tissues: 21 of 29 skin lesions and three of six internal organs); eight of these cats are still alive and six were euthanized. Monoclonal populations were seen in three of five cats that had involvement of internal organs. These findings indicate that feline CL is best considered as a slowly progressive process which may be reactive, but often evolves into a low‐grade indolent lymphoma. Funding: George H. Muller Fund for Research in Dermatology.     

Participation of phosphofructokinase,malate dehydrogenase and isocitrate dehydrogenase in capacitation and acrosome reaction of boar spermatozoa

The aim of this work was to determine the enzymatic activity of phosphofructokinase (PFK), malate dehydrogenase (MDH) and isocitrate dehydrogenase (IDH) in boar spermatozoa and study their participation in bicarbonate‐induced capacitation and follicular fluid‐induced acrosome reaction. Enzymatic activity of these enzymes was determined spectrophotometrically in extracts of boar spermatozoa. Sperm suspensions were incubated in the presence of bicarbonate (40 mM), a well‐known capacitation inducer, or follicular fluid (30%), as an acrosome reaction inducer, and different concentrations of oxoglutarate, oxalomalate and hydroxymalonate, inhibitors of PFK, IDH and MDH, respectively. Capacitation percentages were determined by the fluorescence technique of chlortetracycline (CTC), and true acrosome reaction was determined by trypan blue and differential–interferential contrast, optical microscopy. The activity of PFK in boar spermatozoa enzymatic extracts was 1.70 ± 0.19 U/10¹⁰ spermatozoa, the activity of NAD‐ and NADP‐dependent IDH was 0.111 ± 0.005 U/10¹⁰ and 2.22 ± 0.14 U/10¹⁰ spermatozoa, respectively, and the activity of MDH was 4.24 ± 0.38 U/10¹⁰ spermatozoa. The addition of the specific inhibitors of these enzymes prevented sperm capacitation and decreased sperm motility during capacitation and inhibited the acrosome reaction (AR), without affecting the sperm motility during this process. Our results demonstrate the participation of PFK, IDH and MDH in bicarbonate‐induced capacitation and follicular fluid‐induced acrosome reaction in boar spermatozoa, contributing to elucidate the mechanisms that produce energy necessary for these processes in porcine spermatozoa.     

Effect of feeding different levels of wheat roti on nutrient utilization and blood metabolite profile in semi‐captive Asian elephants (Elephas maximus)

This experiment was conducted to study the effect of different levels of wheat roti (WR) on nutrient utilization and blood metabolites in Asian elephants fed roughages ad libitum. Nine (3 M, 6 F) Asian elephants (14–52 years of age, 1909–3968 kg BW) were used in an experiment based on replicated Latin square design. Animals in each group (n = 3) were assigned to one of the three dietary treatments in a manner that animals in all the three groups were exposed to all the three treatments in three different periods. Each feeding trial comprised 30 days (25 days of adaptation and 5 days collection period). The amount of WR fed to the elephants was 0.18, 0.12 and 0.06% of BW in groups I, II and III, respectively. They were allowed to forage in the nearby forests for 6 h/day and to bathe for 2 h/day. The animals had ad libitum access to cut Rohini (Mallotus philippensis) trees in their night shelter. Intake and apparent digestibility of dry matter (DM), crude protein (CP), gross energy (GE), Ca, P, Fe, Cu and Zn were measured. Feed consumption was not significantly different among the groups. Significant (p < 0.01) decrease in digestibility of DM and GE and blood glucose concentration was observed with decreased level of WR in the diet. Feeding of WR at 0.06% of BW supplied adequate amount of DE, CP, Ca, P, Fe, Cu and Zn to meet requirement for adult maintenance. Feeding of WR in excess of 0.06% of BW supplied DE in excess of requirement, increased blood glucose concentration which may cause obesity and other associated health problems. It was concluded that the amount of WR should be restricted to 0.06% of BW in the diet of captive Asian elephants.