Sensitivity of African swine fever virus (ASFV) to heat, alkalinity and peroxide treatment in presence or absence of porcine plasma.

African swine fever virus (ASFV) is a highly resistant viraemic virus with devastating socio-economic impact. Its present epidemiology in Eastern Europe and Russia warrants increased biosecurity measures in Western Europe. This includes proactive precautions on traffic of pork products within and between areas that are officially free from ASF. Namely, delayed notification of clinical signs or introduction of a low-virulent strain in ASF-free areas could result in presence of ASFV in veterinary inspected pork and pork by-products. The present study evaluated sensitivity of ASFV to physical and chemical processing conditions that can be applied on abattoir collected blood for production of spray dried porcine plasma (SDPP). Standard endpoint dilution assays were used to determine the sensitivity of Vero-cell adapted Lisbon/60 strain ASFV to heat treatment (H) at alkaline conditions (A) with or without peroxide (P). Time (T) dependent inactivation was evaluated in presence or absence of porcine plasma. HAPT-treatment at H = 48 °C, A = pH 10.2 and P = 20.6 or 102.9 mM H2O2 during 10 min (T) inactivated (95LCL) 3.35, respectively, 4.17 log10 TCID50 ASFV/ml plasma. In absence of plasma, 6.99 log-inactivation was reached within 5 min. Implementation of HAPT-treatment on plasma from ASFV-free areas provides an additional safety hurdle for derived blood products in the unlikely event that blood from few undetected infected pigs would contaminate pooled veterinary inspected blood. Such an additional processing step in the production of SDPP is thus a valuable precautionary measure to overcome a potential biosecurity-break that may arise during the high-risk phase between transboundary introduction of ASFV and first notification of the disease.

Sensitivity of porcine epidemic diarrhea virus (PEDV) to pH and heat treatment in the presence or absence of porcine plasma.

Emergence of porcine epidemic diarrhea virus (PEDV) resulted in massive neonatal mortality in the North-American and Asian pork industry. Measures to prevent its geographical spread are of utmost importance to safeguard susceptible porcine populations. The major infection route is direct or indirect faecal-oral contact. Adequate biosafety measures should be in place at all levels of the swine production chain, including feed and feed ingredients. Present study aimed to investigate the sensitivity of PEDV to thermal inactivation at neutral and alkaline pH in presence or absence of porcine plasma. Cell culture medium and porcine plasma at different pH (7.2, 9.2, 10.2) and temperature conditions (4 °C, 40 °C, 44 °C, 48 °C) were inoculated to a final titer of 5.5 log10 TCID50 PEDV/ml, incubated for up to 120 min and the residual infectivity was determined by endpoint dilution assay. Irrespective of presence of plasma, PEDV was not sensitive to pH 7.2-10.2 at 4 °C. At moderate temperatures (≥40 °C), both alkaline pH and presence of plasma potentiated thermal inactivation. Inactivation of 8 log10 TCID50/ml plasma within 30 min (8D value<30 min) by moderate pH and temperature would denote potential industrial processing conditions that ensure safety towards PEDV while limiting denaturation of bioactive components. Virus-spiked plasma required heat treatment of 40 °C and alkalinization to pH 9.2 to achieve 8 log10 reduction within such time. At pH 10.2 and 48 °C, the 8D value was 4.6 min in plasma and 15.2 min in MEM. Here we propose heat-alkalinity-time (HAT) pasteurization as a highly efficient method to inactivate PEDV during industrial processing of porcine plasma.

Voluntary feed intake and leptin sensitivity in ad libitum fed obese ponies following a period of restricted feeding: a pilot study.

The relation between plasma leptin and daily ad libitum roughage intake was evaluated during a 14-day period in eight obese Shetland ponies. When the feeding strategy was changed from maintenance feeding to ad libitum feeding, feed intake increased the first day, decreased the second day and increased again during the following days to reach a constant level after 8 days. Plasma leptin concentration increased during the first 2 days, but remained constant afterwards. Although the same pattern was found in all ponies, the magnitude of the increase in leptin on day 1 and the resulting decrease in feed intake on day 2 differed between ponies. A lower anorectic effect was seen in ponies with higher initial leptin concentration, suggesting the presence of different degrees in leptin sensitivity in obese ponies. High leptin production in a attempt to compensate for the decrease in leptin sensitivity might explain large variations in plasma leptin among obese ponies with similar body condition score. Further research is necessary to clarify whether the reduced leptin sensitivity precedes obesity in equines or vice versa.

Chlamydial infections in Chinese livestock.

The occurrence and impact of chlamydial infections in Western livestock is well documented in the international literature, but less is known aboutthese infections in livestock in the People's Republic of China. China's livestock production and its share in the global market have increased significantly in recent decades. In this review, the relevant English and Chinese literature on the epidemiology of chlamydial infections in Chinese livestock is considered, and biosecurity measures, prophylaxis and treatment of these infections in China's livestock are compared with Western practices. Chlamydial infections are highly prevalent in Chinese livestock and cause important economic losses, as they do in the rest of the world. Surveillance data and diagnostic results of abortion outbreaks in cattle, sheep and goats highlight the importance of virulent chlamydial infections in China's major ruminant species in many of China's provinces, autonomous regions and municipalities. Data from many of China's provincial divisions also indicate the widespread presence of chlamydial infections in industrially reared swine across the country. Less is known about chlamydial infections in yak, buffalo and horses, but available reports indicate a high prevalence in China's populations. In these reports, chlamydiosis was related to abortions in yak and pneumonia in horses. In Western countries, chlamydial infections are principally treated with antibiotics. In China, however, traditional medicine is often used in conjunction with antibiotics or used as an alternative treatment.

Glycine and its N-methylated analogues cause pH-dependent membrane damage to enterotoxigenic Escherichia coli.

The current study first investigates the emulsifying potential of glycine and its N-methylated derivatives N-methylglycine (sarcosine), N,N-dimethylglycine (DMG) and N,N,N-trimethylglycine (betaine) under varying pH conditions. Subsequently, the effect of these test compounds on the membrane integrity of enterotoxigenic Escherichia coli (ETEC) was evaluated. Oil in water emulsions containing each compound show that DMG is a more potent enhancer of emulsification than glycine, sarcosine and betaine under the conditions tested. Flow cytometry was used to investigate whether the emulsifying potential is associated with an effect on ETEC membrane integrity. The bacteria were exposed to each of the test compounds under varying pH conditions and membrane integrity was assessed using the LIVE/DEAD BacLight kit. Results show a membrane deteriorating effect caused by glycine, sarcosine and DMG, but not by betaine. This effect is pH- and time-dependent and has an apparent threshold at pH 9.0. Conventional plate counts confirmed concomitant changes in culturability of the membrane comprised bacteria.

Indirect evidence for microbiota reduction through dietary mannanoligosaccharides in the pigeon, an avian species without functional caeca.

A feeding trial was conducted to evaluate the effects of mannanoligosaccharides (MOS) on nutrient digestibility, intestinal pH, gut morphology and faecal bacteriology of pigeons, as model for birds without functional caeca. Sixteen adult pigeons (Columba livia domestica) were randomly allotted to either an extruded pellet diet with or without 0.4% MOS. After an adaptation period of 24 days, excreta were collected during 4 days. Apparent nutrient digestibility coefficients were determined using total collection method. Further, excreta pH was measured and percentage of uric acid determined. Fresh excreta were cultured for measurement of colony-forming units for Escherichia coli. At the end, the gastrointestinal tract (GIT) was excised and pH measurements performed on the separate GIT sections. Finally, pancreas, liver, gizzard and abdominal fat pad were weighed, and standardised segments of duodenum and jejunum were removed for microscopic measurement of crypt depth, villus height and muscularis thickness. Feed intake and water intake were similar between control diet and MOS diet. Intestinal pH was unaffected by MOS supplementation; however, excreta pH was significantly lower in pigeons on the MOS diet. Although nutrient digestibility was similar between treatments, uric acid content of excreta was significantly higher in the MOS group in relation to the control group. Further, duodenal crypt depth, villus height and muscularis thickness, as well as jejunal muscularis thickness were all significantly reduced by MOS supplementation. No effect of MOS supplementation was seen on the counts of E. coli. Furthermore, despite marked differences on both GIT morphology and uric acid content of excreta, apparent digestibility coefficients, and organ weights, were similar between treatments. It is suggested that the MOS-induced changes on gut morphology and the reduced excreta pH reflect a reduced bacterial challenge in the intestine of pigeons. Supplementation of MOS, therefore, has potential as prebiotic strategy in birds without functional caeca.

The effect of starch gelatinisation degree on intestinal morphology, intestinal pH and bacteriology in pigeons.

The effect of starch gelatinisation degree in extruded feed on intestinal morphology, intestinal pH and faecal bacteriology was investigated in pigeons. Extruded complete pigeon diets would offer the principle advantage of providing equilibrated nutrients and energy, but factors such as starch gelatinisation require investigation before these diets are offered as main dietary items to pigeons. Birds were fed two diets with equal ingredient composition and nutrient content, but with a different degree of starch gelatinisation resulting from altered extrusion processing (high gelatinisation degree (HG) with 73.6% gelatinisation vs. low gelatinisation degree (LG) with 53.1% gelatinisation). Feed intake and weight gain changes were measured weekly. Blood samples were collected at day 28 and analysed for non-esterified fatty acids, lactate dehydrogenase and glucose. The pH values for fresh excreta were measured; thereafter fresh excreta were collected and cultured for measurement of colony-forming units for bacterial classes. At the end, morphological measurements were examined and the pH values throughout the gastrointestinal tract were recorded. Liver, pancreas and abdominal fat were weighed. There was a tendency (p= 0.07) towards higher numbers of Escherichia coli in the excreta of the LG group compared with those in the HG group. No dietary treatment effects were noted on the number of Lactobacillus sp. in the excreta. In proximal parts of the intestine, LG revealed a significantly lower pH than HG. Villus height and crypt depth were not affected by dietary treatment, but the duodenum muscularis thickness, liver weight and pancreas weight were significantly lower in the LG than that in HG group. This trial demonstrated that the lower level of starch gelatinisation degree of extruded feed leads to acidification of the proximal gut and altered gut morphology in pigeons. Hence, extruded pigeon diets should preferably contain low-gelatinised starch instead of high-gelatinised starch. In addition, future research must focus on the effects of starch gelatinisation on the correlation between the intestinal pH, microflora content and intestinal morphology.

Dietary supplementation with dimethylglycine affects broiler performance and plasma metabolites depending on dose and dietary fatty acid profile.

The effect of dietary supplementation with N,N-dimethylglycine sodium salt (Na-DMG) was evaluated in a feeding trial with 1500 1-day-old broiler chicks (Cobb 500). DMG was supplemented at 0, 0.1, 0.2, 0.5 or 1 g Na-DMG/kg feed to a ration with either animal fat (chicken fat) or vegetal fat (soy oil) as main fat source. In the vegetal fat diets, production value was significantly linearly improved by supplementation with DMG up to 11%. Irrespective of dietary fat source, abdominal fat percentage was significantly linearly reduced up to 24% and meat yield tended to increase linearly with DMG level up to 4%. In the vegetal fat groups, DMG significantly lowered abdominal fat pad by up to 38% and tended to increase meat yield up to 6% at the highest dose. Fasted non-esterified fatty acid level significantly decreased with increasing DMG level up to 36% and thiobarbituric acid reactive species (TBARS) decreased with a statistical trend up to 46% at the highest dose. In vegetal fat diets, addition of DMG resulted in significant lower TBARS level by 56% at the highest dose. Finally, a significant quadratic effect on ascites heart index was present in the vegetal fat diets, with a minimal value at 0.5 g Na-DMG/kg. In conclusion, dietary supplementation with DMG may improve technical and slaughter performance, and may reduce oxidative stress and pulmonary hypertension, but the degree of effects is modulated by fatty acid profile of the diet. Herewith, effects are more pronounced in a diet rich in polyunsaturated fatty acids compared with a diet rich in saturated and monounsaturated fatty acids.

Effects of segregation and impact of specific feeding behaviour and additional fruit on voluntary nutrient and energy intake in yellow-shouldered amazons (Amazona barbadensis) when fed a multi-component seed diet ad libitum.

Parrots are commonly fed multi-component seed diets; however, both segregation and feeding behaviour might alter ingredient and nutrient composition of the offered diet. First, the nutritional impact of segregation was assessed as it occurs when multi-component diets are temporarily stored in food containers that are replenished before completely emptied and birds being fed from the upper layer. The most detrimental effect hereof was a vast decrease in mineral supplements, leading to a decrease in Ca:P ratio in the offered food in relation to the formulated diet. Next, caloric distribution shifted towards more EE energy at the expense of NFE energy, as proportion of oilseeds increased and NFE-rich seeds decreased. Next, a feeding trial was performed on six yellow-shouldered amazons (Amazona Barbadensis) in which nutritional impact of parrot-specific feeding behaviour was assessed as well as the influence of additional provision of fruit next to the seed mixture. Profound selective feeding behaviour and dehusking of seeds resulted in a vast increase in energetic density by up to 64% in the ingested fraction in relation to the offered mixture in toto. Furthermore, the already suboptimal Ca:P ratio further deteriorated and caloric distribution shifted by over twofold towards EE energy accompanied with a vast decline in NFE energy, CP energy remaining similar. Finally, provision of fruit next to the seed diet significantly lowered voluntary energy intake from 936 ± 71 to 809 ± 109 kJ ME/kg(0.75)/day, without compromising adequate protein intake. In conclusion, notwithstanding efforts of nutritionists to formulate diets to approximate estimated, species-specific requirements, nutritional composition of the actually consumed fraction of multi-component seed diets can be vastly deteriorated by both animal and management factors. Furthermore, offering of fruit next to a seed-based diet effectively reduces voluntary energy intake and can hence be applied to abate obesity.

Dietary N,N-dimethylglycine supplementation improves nutrient digestibility and attenuates pulmonary hypertension syndrome in broilers.

N,N-dimethylglycine (DMG) is an intermediary metabolite in cellular choline and betaine metabolism. The present trial aimed to evaluate the effect of dietary DMG on nutrient digestibility and development of pulmonary hypertension syndrome in broilers. A total of 64 14-day-old broiler hens (Ross-308) were raised until age 40 days under cold environmental temperature conditions (15 °C) and were fed a high energy feed in order to incite pulmonary hypertension. Birds were randomly assigned to two groups of which each group had eight replicate pens of four birds each. Test diets contained 0 or 167 mg Na-DMG (Taminizer(®) D; Taminco N.V., Ghent, Belgium)/kg feed. N,N-dimethylglycine supplementation resulted in a significant improvement in apparent faecal digestibility of crude protein and nitrogen-free extract. Further, fulminant ascites was numerically lowered by DMG and incidence of pulmonary hypertension decreased significantly from 44.8% in the control group to 14.6% in the DMG group. Finally, fasted plasma level of non-esterified fatty acids (NEFA) was twofold in the control group in relation to the DMG group. In conclusion, these data demonstrate beneficial effects of DMG on digestibility of non-fat fractions, on fat metabolism and on progression towards broiler ascites syndrome.

Effect of N,N-dimethylglycine supplementation in parturition feed for sows on metabolism, nutrient digestibility and reproductive performance.

The current pilot study assessed the influence of N,N-dimethylglycine (DMG) on insulin sensitivity, glucose and fat metabolism, nutrient digestibility and reproductive performance of sows in the peripartal period. At day 105 of gestation, 25 sows were randomly assigned to the control (n = 13) or the DMG group (n = 12). Sows from the DMG group were supplemented with 1 g DMG/kg feed until day 3 of lactation. After an overnight fast 1 day after farrowing, a blood sample of each sow was drawn. The plasma was analyzed for insulin, glucose, fructosamine, leptin, thiobarbituric acid reactive substances (TBARS), ferric reducing ability of plasma (FRAP), non-esterified fatty acids (NEFA) and triglycerides (TG) and an oral glucose tolerance test was performed. A rectal feces sample was collected and the apparent fecal digestibility (AFD) of crude fat (CFAT), crude protein (CP) and nitrogen-free extract (NFE) was calculated after proximate analyses. Finally, a colostrum sample was collected from each sow and analyzed for the presence of DMG. Reproductive performance parameters were recorded. The results showed an improvement in the AFD of CFAT, CP and NFE when DMG was supplemented. This beneficial effect confirms the hypothesis that DMG acts as an emulsifying agent. The improvement in digestibility in the DMG group was accompanied by a numerical increase in plasma TG (P = 0.067). Plasma NEFA concentrations were not different between treatment groups. DMG supplementation neither affected glucose clearance nor influenced plasma insulin, glucose, fructosamine or leptin levels. TBARS and FRAP also remained unaffected, despite previously reported anti-oxidative properties of DMG. Furthermore, no significant impact on reproductive performance could be recorded. In conclusion, DMG supplementation significantly improved nutrient digestibility. Possible beneficial effects on energy metabolism and reproductive performance of sows should be tested when DMG is supplemented for a longer period of time or at a higher dose.

Effect of dilution degree of commercial nectar and provision of fruit on food, energy and nutrient intake in two rainbow lorikeet subspecies.

A feeding trial was performed on adult rainbow lorikeets of two subspecies: six green-naped lorikeets (Trichoglossus haematodus haematodus) and six red-breasted lorikeets (T. haematodus mitchellii). Throughout the entire trial, half of the birds from each subspecies had ad libitum access to water-diluted commercial nectar powder and drinking water only, whereas the other half also received ad libitum apple pieces. During three consecutive 14-d periods, the nectar powder was diluted to a different degree: 1:3 (low), 1:5 (high) and 1:4 (medium) (v:v). Diluting nectar to a higher degree resulted in both subspecies in a decrease in voluntary energy intake. Next, nectar intake significantly decreased when apple was available and apple intake significantly increased when fed higher-diluted nectar. In green-naped lorikeets fed nectar and apple, energy intake was similar between dilution degrees of nectar but was lower compared with feeding only low- or medium-diluted nectar. Whereas, in red-breasted lorikeets, provision of apple next to medium- or high-diluted nectar resulted in higher voluntary energy intake compared with feeding solely nectar of any degree. Overall, protein and thiamine intake as well as Ca:P ratio of the ingested ration were lowest when fed highly diluted nectar and apple. Yet, minimal requirements were still covered. Because energy content of fruit can be higher than liquid diets, in this case medium- or high-diluted nectar, ad libitum provision of fruit as a means to lower voluntary energy intake in lorikeets, for instance in case of obesity, needs to be considered with care.

Apparent nutrient digestibility and excreta quality in African grey parrots fed two pelleted diets based on coarsely or finely ground ingredients.

A feeding trial was performed to study the influence of particle size in extruded parrot pellets on apparent digestibility and excreta consistency and pH. Two test diets were alternately provided to eight African grey parrots according to a 2 x 2 cross-over design. Both diets were similar in nutrient content and ingredient composition but differed in particle size of the composing particles of individual pellets. Apparent digestibility of macronutrients was studied using the total collection method. Next, the appearance of the excreta was studied by calculation of weight-surface ratio of individual excrements as an objective measurement of consistency. Last, excreta pH was measured directly on fresh excrements and on homogenized 10% excreta solutions. Neither apparent digestibility coefficients nor excreta pH values were significantly different in parrots fed the two diets. However, excreta consistency was significantly (p < 0.05) more solid when fed the coarse diet than when fed with the finely ground diet. The results of this study suggest that excreta consistency can be improved through larger particle size, without adverse effects on nutritive value of the diet.