Prediction of digestible protein content of dry extruded dog foods: comparison of methods

The aim of this study was to check the accuracy of laboratory methods to predict the apparent protein digestibility (CPd, %) and digestible protein content (DP, g/kgDM) of dog foods, avoiding the use of experimental animals in digestion trials. Twenty‐eight commercial dry extruded dog foods were tested by three different methodologies: an adaptation of the in vitro incubation method described by Hervera et al. (J Anim Physiol Anim Nutr 2007, 91 : 205) for estimation of digestible energy of commercial dog foods, a modification of the pH drop methodology proposed by Hsu et al. (J Food Sci 1977, 42 : 1269) for protein evaluation of human foods and the Near Infra Red Spectroscopy (NIRS) technology. All the methods assessed revealed very good, similar accuracy in the prediction of DP content either using the in vitro method (r = 0.99; RSD = 6.76; CV = 2.31%), the pH drop method (r = 0.99; RSD = 6.94; CV = 3.02%) or the NIRS (R2 = 0.96; SECV = 10.50) method, although the in vitro digestion method showed the highest accuracy approach of in vivo crude protein apparent digestibility: CPd in vitro (r = 0.81; RSD = 2.01; CV = 2.41%); CPd pH‐drop (r = 0.78; RSD = 2.48; CV = 2.98%) and NIRS (r²cv = 0.53; SECV = 2.37).     

Evaluation of serum allergen-specific IgE for the diagnosis of food adverse reactions in the dog

A new monoclonal enzyme-linked immunoassay (ELISA; CMG IMMUNODOT, Fribourg, Switzerland) measuring food antigen-specific serum IgE was used in an attempt to investigate food allergen-specific IgE in dogs. The serum of eight dogs with clinically proven adverse reactions to specific proteins was tested for beef, cow's milk, pork, lamb, hen's egg, soybean, fish mix (cod/sole), peanut, maize and wheat flour. The control group consisted of three healthy dogs, three dogs with nonallergic skin disease, two dogs with atopy, a cat and a horse. Only three mild positive reactions to beef, lamb and peanut, respectively, were found in this study; the sera were from two control dogs with the clinical diagnosis of dermatophytosis and atopy. None of the animals with confirmed food adverse reactions showed positive reactions. This study indicates that the diagnosis of food adverse reactions in the dog by measuring allergen-specific IgE with the used mononuclear ELISA is unreliable.     

Availability, brands, labelling and salmonella contamination of raw pet food in the minneapolis/st. Paul area

The purpose of the study was to characterize the commercially available raw meat pet food diets in the Minneapolis/St. Paul area by (i) determining the number and types of available diets; (ii) assessing pet food stores and brand labels for the provision of precautionary statements regarding the risk of foodborne illness from raw meat; (ii) assessing the labels for Food and Drug Administration (FDA)/American Association of Feed Control Officials (AAFCO) required content and nutrient-related information; and (iv) culturing purchased diets for the presence of Salmonella. Sixty raw meat diets were purchased, representing 11 different brands from eight different stores. Diets were readily available in the form of raw-frozen, dehydrated or freeze-dried varieties from different protein sources, such as lamb, beef, chicken or duck. All stores promoted raw meat diets; however, none provided foodborne illness warnings. Brands varied greatly in their precautionary statements; none of the diets underwent feeding trials; and nutritional adequacy substantiation was through formulation only. The first five ingredients tended to consist of meat, organ meat (by-products), vegetables, grains and ground bones. Currently, it is required that pet foods have an AAFCO nutritional adequacy statement and provide a guaranteed analysis table. Three brands did not meet these FDA requirements. Thirty-one (51.7%) of the 60 raw meat diets underwent some degree of processing including dehydration, freeze-drying or high-pressure pasteurization. Four of the 60 raw diets (7%) tested positive for Salmonella. Analysis of raw meat pet food labels indicated a lack of foodborne illness warnings. Based on these findings, we recommend that warning statements similar to those required by the United States Department of Agriculture and placed on labels of raw meat intended for human consumption be provided on the labels of raw meat pet food diets.     

2006~2007年中国饲料及饲料原料霉菌毒素污染调查报告

作者评估了2006年和2007年上半年我国饲料原料和全价饲料中6种主要霉菌毒素的检出水平和分布特点,得出如下结论：①我国饲料原料和全价饲料中超标率和检出水平最高的霉菌毒素是玉米赤霉烯酮、烟曲霉毒素和呕吐毒素（田间型毒素）。人们一直重视的黄曲霉毒素（仓储型毒素）超标率和检出水平均比较低;②饲料原料和全价饲料中多种霉菌毒素往往同时存在,由于不同毒素间具有协同作用,将加剧毒素的毒性作用和中毒症状的严重程度;③副产品原料,特别是玉米副产品如干燥酒糟（DDGS）和玉米蛋白粉中霉菌毒素的污染非常严重;④与单一能量和蛋白质饲料原料相比,全价饲料中的多种霉菌毒素的检出水平才是相对比较准确和客观的控制指标;⑤控制霉菌毒素危害的最经济有效的措施是控制原料质量和使用有效的霉菌毒素吸附剂。     

ELISA testing for common food antigens in four dry dog foods used in dietary elimination trials

This study evaluated four over the counter venison dry dog foods available from one on-line retail vendor for potential contamination with common known food allergens: soy, poultry or beef. An amplified, double sandwich type enzyme linked immunosorbent assay (ELISA) test of soy, poultry and beef proteins were performed by an independent accredited food laboratory. The ELISA test for poultry protein was found to be unreliable when testing in dry dog foods because false negatives occurred. ELISA testing of control diets for both soy and beef proteins performed as expected and could be useful in antigen testing in dry dog foods. Three of the four over the counter (OTC) venison canine dry foods with no soy products named in the ingredient list were ELISA positive for soy; additionally one OTC diet tested positive for beef protein with no beef products listed as an ingredient list. One OTC venison diet was not found to be positive for soy, poultry or beef proteins. However, none of the four OTC venison diets could be considered suitable for a diagnostic elimination trial as they all contained common pet food proteins, some of which were readily identifiable on the label and some that were only detected by ELISA. Therefore, if the four OTC venison products selected in this study are representative of OTC products in general, then the use of OTC venison dry dog foods should not be used during elimination trials in suspected food allergy patients.     

Prediction of digestible energy content of extruded dog food by in vitro analyses

The aim of this study was to develop a simple and reproducible in vitro method for predicting the apparent energy digestibility of dry extruded dog foods. The proposed method is based on the two-step multienzymatic incubation assay described by [Boisen, S., 1991: In Vitro Digestion for Pigs and Poultry, M. F. Fuller (ed.). CAB International, Wallingford, 135-146], with some modifications adapted to dogs' digestion characteristics. The method consisted in two consecutives incubations, first one during 2 h with pepsin (10 mg/g of food sample) in acid pH and second one during 4 h with pancreatin (100 mg/g of food sample). The undigested residue obtained was collected in a filtration unit and then dried and ashed. The in vitro percentage of organic matter disappearance (in vitro dOM) of 54 dry extruded commercial dog foods was determined and used as predictor of the in vivo apparent organic matter (in vivo dOM) and energy digestibility (in vivo dE) and digestible energy (DE) content. There was a close linear relationship between the in vivo dOM and dE [r(2) = 0.95, residual standard deviation (RSD) = 1.05 and coefficient of variation (CV) = 1.2%] and also between the in vitro and in vivo dOM (r(2) = 0.92, RSD = 1.38 and CV = 1.6%), even if the in vitro dOM overestimated on average by 4% the in vivo dOM. When the in vitro dOM was used to predict the in vivo dE, the relationship between both variables was defined by the equation: in vivo dE (%) = -2.45 + 0.98 +/- 0.04x in vitro dOM (%), (r(2) = 0.92, RSD = 1.25 and CV = 1.5%). In addition, a close relationship between the in vivo and predicted DE (estimated dE x gross energy determined calorimetrically) was found (r(2) = 0.97, RSD = 0.26 and CV = 1.4%). The accuracy of DE content prediction using the proposed in vitro method was higher than that obtained when the DE content of the same set of samples was predicted by the equation proposed by the NRC (1985) (r(2) = 0.76, CV = 4.5%) and also slightly higher than that obtained when using the equation proposed by NRC (2006) (r(2) = 0.95, CV = 2.1%).     

Effects of the ruminal comminution rate and microbial contamination of particles on accuracy of in situ estimates of ruminal degradability and intestinal digestibility of feedstuffs

Effects of considering the comminution rate (k_c) and the correction of microbial contamination (using ¹⁵N techniques) of particles in the rumen on estimates of ruminally undegraded fractions and their intestinal digestibility were examined generating composite samples (from rumen‐incubated residues) representative of the undegraded feed rumen outflow. The study used sunflower meal (SFM) and Italian ryegrass hay (RGH) and three rumen and duodenum cannulated wethers fed with a 40:60 RGH to concentrate diet (75 g DM/kgBW^0.75). Transit studies up to the duodenum with Yb‐SFM and Eu‐RGH marked samples showed higher k_c values (/h) in SFM than in RGH (0.577 vs. 0.0892, p = 0.034), whereas similar values occurred for the rumen passage rate (k_p). Estimates of ruminally undegraded and intestinal digestibility of all tested fractions decreased when k_c was considered and also applying microbial correction. Thus, microbial uncorrected k_p‐based proportions of intestinal digested undegraded crude protein overestimated those corrected and k_c?k_p‐based by 39% in SFM (0.146 vs. 0.105) and 761% in RGH (0.373 vs. 0.0433). Results show that both k_c and microbial contamination correction should be considered to obtain accurate in situ estimates in grasses, whereas in protein concentrates not considering k_c is an important source of error.     

Microbial contamination of the anterior chamber during cataract phacoemulsification and intraocular lens implantation in dogs

The objective of this study was to determine the frequency of intraoperative contamination of the anterior chamber with viable microorganisms during cataract phacoemulsification and intraocular lens implantation, and to evaluate the relationship of contaminant microorganisms to patients' extraocular and nasal cavity floras. Also, the impact of various aspects of the patient history and phacoemulsification procedure on the incidence of positive postoperative anterior chamber cultures was investigated. Twenty-two eyes from 13 dogs presented for elective cataract phacoemulsification and intraocular lens implantation were studied. Preoperatively, microbiologic samples of the conjunctiva, eyelid margins, nares, and rostral nasal cavity were collected. Postoperatively, anterior chamber fluid was aspirated. Samples were submitted for aerobic/anaerobic bacteriologic culture and antimicrobial susceptibility, Mycoplasma culture, and fungal culture. Anterior chamber aspirates collected at the conclusion of surgery were culture positive for at least one organism in 22.7% of eyes. Three aerobic bacteria and three fungi were isolated from the anterior chamber aspirates. Two fungi and one bacterium isolated from the anterior chamber were typed identically, and the bacterium had a similar antibiogram to organisms recovered from the patient's conjunctiva and eyelid margin. No statistically significant difference in contamination frequency was found for the investigated patient and surgical variables. We conclude that intraoperative contamination of the anterior chamber with viable bacterial and fungal organisms is a common occurrence in canine patients undergoing cataract phacoemulsification and intraocular lens implantation, and the external ocular flora is a likely source of some of these contaminating microorganisms. This contamination is independent of the patient and surgical variables investigated.     

兰州市肉与肉制品微生物污染状况的研究

目的：了解兰州市定点屠宰场生猪屠宰加工产品的卫生质量以及肉与肉制品致病菌污染状况。方法：2003-2006年,应用国标法对2个定点生猪屠宰场300份生猪胴体样品进行了菌落总数、大肠菌群和沙门氏菌检验;采用随机采样法对兰州市不同酒店和超市的生猪肉、牛肉、羊肉、禽肉100份,熟肉制品1 200份进行了常见致病菌的检验。结果：生猪胴体体表和肉样沙门氏菌检出率分别为7.5%和14%;生肉检出致病菌阳性20份,总检出率为20%;熟肉制品检出致病菌阳性39份,阳性率为3.25%。生肉中以猪肉致病菌阳性率最高,为20%。结论：兰州市生猪胴体的卫生质量急需提高,屠宰生产加工水平亟待改善。     

Pseudomonas fluorescens contamination of a feline packed red blood cell unit and studies of canine units

Background: While screening programs have reduced the risk of infectious disease transmission by donors in human and veterinary blood banking, bacterial contamination of blood products has emerged as a major complication in human medicine. Objectives: To describe a Pseudomonas fluorescens (Pf)‐contaminated feline packed RBC (pRBC) unit and experimentally investigate Pf‐contaminated canine pRBCs. Methods: Canine pRBCs were inoculated with Pf‐rich pRBCs from the sentinel feline unit and stored at 4°C or 20°C for 72 hours. Aliquots from the pRBCs were serially evaluated by microscopy, culture, and a eubacterial 16S rRNA real‐time PCR assay. Results: One Pf‐contaminated feline unit turned black after 22 days of storage and was removed from the blood bank; a source was not found, and no other contaminated units were identified. Canine pRBCs spiked with 5 or 25 μL of the sentinel unit became culture‐ and/or 16S PCR‐positive at ≥8 hours at 20°C and 48 hours at 4°C and developed a color change at ≥24 hours. Sensitivity studies indicated that without incubation, inoculation of ≥100 μL Pf‐rich pRBCs was necessary for a positive 16S PCR test result. Conclusions: P. fluorescens grows in stored pRBCs slowly at 4°C and rapidly at 20°C. Screening of blood products for color change, estimating bacterial concentration with microscopy, and 16S PCR testing are simple and fast ways to detect bacteria in stored blood. Aseptic collection, temperature‐controlled storage, and regular visual monitoring of stored units is recommended. Discolored units should not be transfused, but examined for bacterial contamination or other blood product quality problems.     

Plasma lipoprotein concentrations in the dog: the effects of gender,age, breed and diet

Earlier studies of canine lipoprotein metabolism have frequently not taken into account such variables as age, gender, lifestyle or feeding status. In the last years, many changes to lifestyle and feeding of dogs have occurred. In this study, C-tot, C-HDL, C-LDL, triglycerides and lipoprotein fractions were determined in 251 healthy dogs by means of enzymatic methods and through the electrophoretic technique. All data were analysed by multifactor anova test to determine which factors (age, gender, breed and diet) have a statistically significant effect (p < 0.05) on the determined parameter and subsequently Bonferroni’s test was applied where necessary. Gender, age, breed and diet can significantly affect lipid metabolism, in particular lipoproteins involved in cholesterol plasma transport; on the contrary, triglycerides are not influenced by the same factors. The most important observation about age is the high level of C-LDL in puppies under 1 year of age. The highest cholesterol concentrations are found in Rottweiler but high values of plasma cholesterol are found also in Pyrenees Mountain dog and a great level of C-LDL in Labrador. Diet has shown a great influence on lipidic metabolism: dogs fed with different high-quality dry foods had significant differences in plasma cholesterol values (C-tot, C-HDL, C-LDL,), in particular, dogs fed with a diet rich in fish and fish-by-products have shown the lowest levels of C-tot, C-HDL and C-LDL.     

Processing traits and digestibility of extruded dog foods with soy protein concentrate

Soya bean protein concentrate (SPC) with two particle sizes were evaluated on extrusion parameters, kibble formation, digestibility and palatability of dog foods. Eight diets were extruded: PBM—control diet based on poultry by‐product meal (PBM); GM—a diet in which corn gluten meal (GM) replaced 45% of the diet protein; cSPC15%, cSPC30% and cSPC45%—diets in which SPC of coarse particle size (600 μm) replaced 15%, 30% and 45% of the diet protein; and sSPC15%, sSPC30% and sSPC45%—diets in which SPC of small particle size (200 μm) replaced 15%, 30% and 45% of the diet protein. The digestibility of nutrients was evaluated for the PBM, GM, cSPC45% and sSPC45% diets, using six dogs per food. The PBM, GM and cSPC45% diets were compared for palatability. Data were submitted for analysis of variance, and the means were compared by polynomial contrasts or Tukey's test (p < .05). The cSPC increased the specific mechanical energy (SME) application, extrusion temperature and pressure linearly, resulting in lower kibble density and higher expansion and starch gelatinization (SG) (p < .01). When comparing the PBM, GM, cSPC45% and sSPC45% diets, higher SME, extrusion temperature and pressure, SG and kibble expansion were verified for the cSPC45% diet (p < .05). The DM, fat and crude protein digestibility were similar among diets. Faecal pH, ammonia and lactate did not differ, demonstrating that the removal of oligosaccharides and soluble non‐starch polysaccharides of SPC produces an ingredient with mostly non‐fermentable fibre. Dogs preferred the PBM to the GM diet (p < .05), but consumed the PBM and cSPC45% foods equally. In conclusion, SPC exhibited good extrusion functionality, favouring kibble expansion and SG, with high digestibility, similar to that of PBM. The removal of soluble compounds from soya beans resulted in an ingredient with low fermentable fibre content, which did not alter faecal formation or characteristics.     

Prediction of chemical composition and peroxide value in unground pet foods by near‐infrared spectroscopy

The massive development of the pet food industry in recent years has lead to the formulation of hundreds of canine and feline complete extruded foods with the objective of meeting both the needs of the animals and numerous demands from pet owners. In the meantime, highly variable raw material compositions and the industry's new production techniques oblige manufacturers to monitor all phases of the extrusion process closely in order to ensure the targeted composition and quality of the products. This study aimed at evaluating the potential of infrared technology (visible and near‐infrared spectrophotometer; 570–1842 nm) in predicting the chemical composition and peroxide value (PV) of unground commercial extruded dog foods. Six hundred and forty‐nine commercial extruded dog foods were collected. For each product, an unground aliquot was analysed by infrared instrument while a second aliquot was sent to a laboratory for proximate analysis and PV quantification. The wide range of extruded dog food typologies included in the study was responsible for the wide variability observed within each nutritional trait, especially crude fibre and ash. The mean value of the 208 pet foods sampled for PV quantification was 17.49 mEq O₂/kg fat (min 2.2 and max 94.10 mEq O₂/kg fat). The coefficients of determination in cross‐validation of NIRS prediction models were 0.77, 0.97, 0.83, 0.86, 0.78 and 0.94 for moisture, crude protein, crude fat, crude fibre, ash and nitrogen‐free extract (NFE) respectively. PV prediction was less precise, as demonstrated by the coefficient of determination in cross‐validation (0.66). The results demonstrated the potential of NIRS in predicting chemical composition in unground samples, with lower accuracy for moisture and ash, while PV prediction models suggest use for screening purposes only.     

Cytologic interpretation of canine cerebrospinal fluid samples with low total nucleated cell concentration,with and without blood contamination

Background: Cerebrospinal fluid (CSF) is potentially altered by iatrogenic blood contamination at the time of sampling due to the addition of blood‐associated leukocytes and protein. Objectives: The objective of this study was to assess whether protein concentration, neutrophil percentage, and the presence of activated macrophages, reactive lymphocytes, or eosinophils in CSF samples with low total nucleated cell concentration (TNCC) are affected by blood contamination or associated with central nervous system (CNS) disease. Methods: Case records from the Royal Veterinary College Diagnostic Laboratory were searched retrospectively for dogs with CSF having ≤5 TNCC/μL. TNCC, RBC, and protein concentrations; neutrophil percentage; and the presence of activated macrophages, reactive lymphocytes, and eosinophils were recorded. Results of magnetic resonance imaging (MRI) also were recorded as a marker of CNS disease. Results: Of 906 cases evaluated, 106 (12%) had blood contamination (>500 RBCs/μL) in CSF. Protein concentration and neutrophil percentage were significantly higher and the presence of eosinophils was more likely in blood contaminated vs noncontaminated samples. Non‐blood‐contaminated samples with activated macrophages or reactive lymphocytes had higher protein concentrations and neutrophil percentages, and those with activated macrophages were more likely to have a positive finding on MRI. Conclusions: Protein concentration, neutrophil percentage, and the presence of eosinophils are significantly affected by blood contamination in canine CSF having low TNCC. Activated macrophages and reactive lymphocytes are not affected by blood contamination, however, and may be useful in identifying dogs with CNS abnormalities.     

Quantitation of serum phospholipase A2 by enzyme-diffusion in lecithin agar gels. A comparative study in man and animals

A sensitive gel-diffusion assay for determination of phospholipase A₂ was developed. PLA₂ standards, serum, faecal and pancreas homogenate samples with PLA₂-activity were allowed to diffuse from wells into agar-gels containing lecithin-membranes. The turbidity cleared radially upon PLA₂-activity. The diameters of the cleared zones showed a linear relationship with the log of the enzyme concentration. Serum samples resulted in some turbidity within the cleared zones. This interference originating from serum lipoproteins could be abolished by hydrophobic absorption. The gel-diffusion method was compared with two other methods for PLA₂, titrimetric and radiometric techniques. Analysis on 37 human patients with acute pancreatitis showed close interrelationship between these methods. The phospholipase A₂ activity in sera from man, the dog, the horse, the cow, the pig and the cat were almost equal, but much less than in the albino rat. No significant differences between PLA₂ activities in pancreatic samples were obtained in different animal species. Of the faecal samples, the cow had the lowest PLA₂ activity. Dogs suffering from pancreatic degenerative atrophy (PDA), had significantly reduced PLA₂ activity both in their pancreas and faeces but not in serum.     

紧实度对青贮玉米有氧稳定期发酵品质、微生物数量的效应研究

本研究旨在分析不同紧实度对全株玉米青贮有氧暴露期间发酵品质和微生物变化的影响,为生产实践选择适宜的青贮紧实度提供参考。以新饲玉10号青贮玉米为材料,发酵装料密度设计为5个梯度(350,400,500,600,700 kg/m³),发酵期为50 d,检测开窖后各紧实度处理第12,24,36,60,108 h青贮发酵品质和主要微生物的变化,并用多通道温度记录仪监测温度变化。结果表明,有氧暴露时间与紧实度的交互作用对pH、乳酸、乙酸和氨态氮含量以及乳酸菌、霉菌、酵母菌和好氧细菌数量产生极显著影响(P<0.01)。开窖108 h后,紧实度为600 kg/m³青贮饲料的乳酸菌数量最多,达到8.17 lg cfu/g FW,其pH值和氨态氮含量最低,霉菌和酵母菌数量最少,分别为5.38和7.72 lg cfu/g FW;且紧实度600 kg/m³有氧暴露后稳定的时间显著高于其他处理(P<0.05),达到100 h。通过综合评价,在有氧暴露后紧实度为600 kg/m³的全株玉米青贮发酵品质及有氧稳定性最好,建议600 kg/m³为全株玉米最佳青贮紧实度。     

Evaluation of serum iso-amylase in the normal dog using an improved electrophoretic technique

Six iso-amylase fractions are known to exist in human serum, three originating from the salivary glands and three from the pancreas. Although it is known that a different number and source of iso-amylase fractions occur in the dog, the routine detection of all the canine iso-amylase fractions has not been previously established. Earlier methods detected either two iso-amylase fractions or, in a proportion of cases, four fractions. A method is described which detects four iso-amylase fractions in 95% of normal canine serum samples. Trials have shown the method is reproducible and that freezing at -40°C has no effect on iso-amylase activity. The normal values for iso-amylase are recorded in 18 normal dogs.