James S. Cullor

Toxoplasma in Animals, Food, and Humans: An Old Parasite of New Concern

All hosts, including humans, can be infected by any one of the three forms of the parasite Toxoplasma gondii that correspond to three morphological stages: tachyzoite, bradyzoite, and sporozoite form. Felids are definitive hosts for T. gondii, which is an intracellular pathogen that infects a wide range of warm-blooded intermediate hosts. Toxoplasmosis is a disease where the interest of the diverse medical and veterinary specialties converge. Awareness needs to be increased that toxoplasmosis can induce clinical disease not only in immunocompromised patients or through congenital infections, but also in healthy patients. This is a review article that aims at illustrating why toxoplasmosis should be regarded a veterinary public health issue and how veterinary practitioners can contribute in controlling the infection.

Enterotoxin production by Staphylococcus aureus isolated from mastitic cows

Staphylococcus aureus is an important cause of mastitis in cows. The ability of S. aureus strains to produce one or more enterotoxins in milk and dairy products is linked to staphylococcal food poisoning. To determine whether staphylococci causing bovine mastitis could cause human foodborne intoxication, the production of staphylococcal enterotoxins A through D (SEA, SEB, SEC, and SED) by 160 S. aureus isolates was evaluated with the use of a reverse passive latex agglutination enterotoxin kit. All S. aureus strains were isolated over a 9-month period from 2,343 routine submissions of a composite quarter collection of individual mastitic cows at 18 dairy farms in the San Joaquin Valley in California. Prior to enterotoxin detection, isolates were grown by a method that enhances the in vitro synthesis of enterotoxin. Twenty-two of 160 S. aureus isolates produced enterotoxin. Seven produced SEC, 12 produced SED, and 3 produced both SEC and SED. None of the isolates produced SEA or SEB.

Quantitative microbial risk assessment for Staphylococcus aureus and Staphylococcus enterotoxin A in raw milk.

A quantitative microbial risk assessment was constructed to determine consumer risk from Staphylococcus aureus and staphylococcal enterotoxin in raw milk. A Monte Carlo simulation model was developed to assess the risk from raw milk consumption using data on levels of S. aureus in milk collected by the University of California-Davis Dairy Food Safety Laboratory from 2,336 California dairies from 2005 to 2008 and using U.S. milk consumption data from the National Health and Nutrition Examination Survey of 2003 and 2004. Four modules were constructed to simulate pathogen growth and staphylococcal enterotoxin A production scenarios to quantify consumer risk levels under various time and temperature storage conditions. The three growth modules predicted that S. aureus levels could surpass the 10(5) CFU/ml level of concern at the 99.9th or 99.99th percentile of servings and therefore may represent a potential consumer risk. Results obtained from the staphylococcal enterotoxin A production module predicted that exposure at the 99.99th percentile could represent a dose capable of eliciting staphylococcal enterotoxin intoxication in all consumer age groups. This study illustrates the utility of quantitative microbial risk assessments for identifying potential food safety issues.

Effect of selected dairy starter cultures on microbiological, chemical and sensory characteristics of swine and venison (Dama dama) nitrite-free dry-cured sausages

The aim of this study was the evaluation of selected lactic acid bacteria (LAB) starter culture of dairy origin in the production of nitrite-free low-acid fermented venison (Dama dama) sausage (salame di daino) produced in a small-scale plant in Umbria (Italy), and their effect on microbiological, physico-chemical and sensorial properties of the products. Salame di daino was obtained with two different processes: with and without the addition of selected LAB starter cultures. Microbial counts of Enterobacteriaceae, coliform organisms and Pseudomonas spp. were lower in salami made with the addition of starter cultures. Staphylococcus aureus, Salmonella spp, and Listeria monocytogenes after the first week of ripening were only detected from control salami. Control salami were paler and harder, whereas those made with the addition of starter cultures were slightly saltier, juicier and in general more acceptable. Selected dairy-origin starter (SDS) cultures did prevent the growth of both indicators of food safety and of process hygiene and increased the acceptability of full-ripened salami.

Efficacy of Ultraviolet (UV-C) Light in a Thin-Film Turbulent Flow for the Reduction of Milkborne Pathogens.

Nonthermal technologies are being investigated as viable alternatives to, or supplemental utilization, with thermal pasteurization in the food-processing industry. In this study, the effect of ultraviolet (UV)-C light on the inactivation of seven milkborne pathogens (Listeria monocytogenes, Serratia marcescens, Salmonella Senftenberg, Yersinia enterocolitica, Aeromonas hydrophila, Escherichia coli, and Staphylococcus aureus) was evaluated. The pathogens were suspended in ultra-high-temperature whole milk and treated at UV doses between 0 and 5000 J/L at a flow rate of 4300 L/h in a thin-film turbulent flow-through pilot system. Of the seven milkborne pathogens tested, L. monocytogenes was the most UV resistant, requiring 2000 J/L of UV-C exposure to reach a 5-log reduction. The most sensitive bacterium was S. aureus, requiring only 1450 J/L to reach a 5-log reduction. This study demonstrated that the survival curves were nonlinear. Sigmoidal inactivation curves were observed for all tested bacterial strains. Nonlinear modeling of the inactivation data was a better fit than the traditional log-linear approach. Results obtained from this study indicate that UV illumination has the potential to be used as a nonthermal method to reduce microorganism populations in milk.

Effects of UV irradiation in a continuous turbulent flow UV reactor on microbiological and sensory characteristics of cow's milk.

The dairy industry under current pasteurization conditions (15 s at 72°C) and sanitary standards achieves a safe product with excellent quality. In an ever-competitive market there is still a need to improve product quality and extend shelf life of dairy products to increase competitiveness and open up new markets. In an attempt to test the effect of UV irradiation on microbiota of fluid milk, a continuous flow UV system at 254 nm was used to treat 3.5 and 2% fat milk at two UV doses (880 and 1,760 J liter(-1)). Milk was obtained from three processors, and two lots from each processor were assessed. To assess the impact on the most descriptive native microbiota in pasteurized milk after UV illumination, the product was held at two storage temperatures (4 and 7°C) and tested weekly for 5 weeks for aerobic plate counts (psychrotrophic and mesophilic bacteria), laboratory pasteurization counts, aerobic sporeformers, coliform organisms, and titratable acidity. Microbial counts for all tested microorganisms were lower in UV-treated milk when compared with control throughout storage at 4 and 7°C in both 3.5 and 2% fat milk. Sensory analysis indicated that there is a sensory defect associated with UV treatment at the wavelength used.

Differential levels of mRNA transcripts encoding immunologic mediators in mammary gland secretions from dairy cows with subclinical environmental Streptococci infections.

Dry-off, and the period around parturition, are associated with increased susceptibility to intramammary infections in dairy cows. The immunological profiles of mammary gland secretions during these periods are not well described. The objective of the present study was to better characterize association(s) between chronic subclinical Environmental Streptococci infections at dry-off and relative levels of mRNA transcripts encoding multiple immunologic mediators present in cells derived from mammary gland secretions at dry-off and continuing through parturition. The chronic subclinical bacterial infections in the present study were characterized by multiple isolations of Streptococcus species and elevated SSC for a minimum of three weeks prior to dry-off. The majority of differences between principal and control quarters were identified at dry-off. Transcript levels of IL-17, IL2Rα and iNOS were increased while pro-inflammatory cytokine IL-6, and the regulatory cytokine IL-10, were reduced. Following antibiotic treatment of mammary glands, IL-17 transcripts remained elevated over the course of the study, indicative of a persistent insult. IL-4 transcript levels were modestly elevated at 7 days following dry-off and significantly elevated at 14 days, consistent with activated T(H)1 and T(H)2 lymphocytes in the principal quarters, respectively. From a temporal perspective, transcript levels of IL-8 decreased in all animals through the dry-off period animals and returned to pre-dry-off levels at parturition; levels of iNOS peaked at parturition. Five of the six principal cows experienced recurrent bacterial mastitis during the subsequent lactation; four were in the same quarter as was initially infected with Streptococcus and three of these four were due to coliforms. Taken together, this apparent chronic susceptibility of select mammary glands to bacterial infection would suggest a physiologic and/or immunologic dysfunction. Identification of factor(s) that contribute to the predisposition of mammary glands to developing mastitis should facilitate development of new control strategies.

Synergistic Effects of Lactic Acid and Sodium Dodecyl Sulfate to Decontaminate Escherichia coli O157:H7 on Cattle Hide Sections

The objective of this study was to investigate the antibacterial properties of chitosan acetate (CA), sodium dodecyl sulfate (SDS), lactic acid (LA) and their synergism when combined against a nontoxigenic strain of Escherichia coli O157:H7. Treatments that significantly reduced the concentration of E. coli O157:H7 in vitro by more than two logs were further investigated using a cattle hide decontamination model. In vitro treatments included CA (1% chitosan in 1% acetic acid vol/vol), SDS (1% vol/vol), SDS (2% vol/vol), LA (1% vol/vol), CA-SDS combination (1% chitosan in 1% acetic acid vol/vol mixed with 1% SDS vol/vol), and LA-SDS combination in two different concentrations (1% LA mixed with 1% SDS vol/vol, and 1% LA mixed with 2% SDS vol/vol). Butterfields Phosphate Buffer water was used as a control. The antibacterial effect of 1% CA solution alone and in combination with 1% SDS in vitro resulted in a 1.8 and 1.7 log colony-forming units (CFU)/mL reduction, respectively (p<0.05). Only 1% LA, 1% SDS, 2% SDS and their combinations resulted in a >2 log reduction in E. coli O157:H7. On hide sections, both 1% LA-1% SDS and 1% LA-2% SDS combinations significantly (p<0.05) reduced E. coli O157:H7 concentration by 4.6 and 4.7 log CFU/ cm(2) greater than the control, respectively. There was no significant difference in the antibacterial effect of 1% LA compared to the control, 2% SDS compared to the control, or 1% LA compared to 2% SDS. Hence, the antibacterial efficacy of 1% LA against E. coli O157:H7 on hide sections was significantly enhanced when combined with 1% SDS. Results of this study support the use of low concentration LA-SDS combination as a hide wash to reduce the risk of E. coli O157:H7 contamination.

Biological markers of neonatal calf performance: The relationship of insulin-like growth factor-I, zinc, and copper to poor neonatal growth

Raising a heifer calf to reproductive age represents an enormous cost to the producer. Poor neonatal growth exacerbates the costs incurred for rearing, and use of blood variables that may be associated with poorly growing calves may offer predictive value for growth and performance. Thus, the principal objective of the present study was to describe changes in serum IGF-I, zinc, and copper from birth to 90 d in Holstein calves, while accounting for sex and twin status, in poorly growing calves and calves growing well. A second objective was to test the hypothesis that an association exists between these serum variables and morphometric indicators of growth. Measurements of BW, length, and height were recorded at birth and at 30, 60, and 90 d of age. Jugular blood (12 mL) was collected from each calf on d 1 to determine serum total protein, serum IgG, packed cell volume, serum zinc, serum copper, serum IGF-I, and CD18 genotype for bovine leukocyte adhesion deficiency; serum zinc, serum copper, and serum IGF-I (predictor variables) were also determined for each calf on d 2 through 10 and on d 30, 60, and 90. Stepwise multiple regression and logistic regression analyses were used to examine the relationships between the predictor variables and the dependent variables (BW, height, and length at d 30, 60, and 90 of life). Birth weight, sex, serum IGF-I (at all ages), serum copper, and the serum copper-to-zinc ratio were associated, to varying degrees, with the dependent growth variables. Birth weight was consistently the dominant predictor. In conclusion, these results suggest that lighter birth weight, reduced serum IGF-I, and inflammation may be important causes of poor growth in neonatal Holstein dairy calves.

Short communication: Attempts to identify Clostridium botulinum toxin in milk from three experimentally intoxicated Holstein cows

Three adult lactating Holstein cows were injected in the subcutaneous abdominal vein with 175 ng/kg of body weight of Clostridium botulinum type C toxin (451 cow median toxic doses) to determine if this botulinum toxin crosses the blood-milk barrier. Whole blood (in sodium heparin) and clotted blood serum samples were taken at 0 min, 10 min, and 3, 6, 9, and 12 h postinoculation. Milk samples were taken at 0 min and at 3, 6, 9 and 12 h postinoculation. All samples were tested for the presence of the toxin using the mouse bioassay and immunostick ELISA test. The immunostick ELISA identified the toxin in whole blood and the mouse bioassay identified the toxin in serum at all times examined in all 3 animals. Toxin was not identified by either detection method in milk samples collected from the 3 animals. From these results, it appears that Clostridium botulinum type C toxin does not cross from the blood to the milk in detectable concentrations.