S. Hussein

Toxicity, metabolism, and impact of mycotoxins on humans and animals.

The worldwide contamination of foods and feeds with mycotoxins is a significant problem. Mycotoxins are secondary metabolites of molds that have adverse effects on humans, animals, and crops that result in illnesses and economic losses. Aflatoxins, ochratoxins, trichothecenes, zearelenone, fumonisins, tremorgenic toxins, and ergot alkaloids are the mycotoxins of greatest agro-economic importance. Some molds are capable of producing more than one mycotoxin and some mycotoxins are produced by more than one fungal species. Often more than one mycotoxin is found on a contaminated substrate. Factors influencing the presence of mycotoxins in foods or feeds include environmental conditions related to storage that can be controlled. Other extrinsic factors such as climate or intrinsic factors such as fungal strain specificity, strain variation, and instability of toxigenic properties are more difficult to control. Mycotoxins have various acute and chronic effects on humans and animals (especially monogastrics) depending on species and susceptibility of an animal within a species. Ruminants have, however, generally been more resistant to the adverse effects of mycotoxins. This is because the rumen microbiota is capable of degrading mycotoxins. The economic impact of mycotoxins include loss of human and animal life, increased health care and veterinary care costs, reduced livestock production, disposal of contaminated foods and feeds, and investment in research and applications to reduce severity of the mycotoxin problem. Although efforts have continued internationally to set guidelines to control mycotoxins, practical measures have not been adequately implemented.

Vegetation Change After 65 Years of Grazing and Grazing Exclusion

Abstract The Nevada Plots exclosure system was constructed in 1937 following passage of the Taylor Grazing Act to assess long-term effects of livestock grazing on Nevada rangelands. A comparison of vegetation characteristics inside and outside exclosures was conducted during 2001 and 2002 at 16 sites. Data analysis was performed with a paired t test. Out of 238 cover and density comparisons between inside and outside exclosures at each site, 34 (14% of total) were different (P < 0.05). Generally, where differences occurred, basal and canopy cover were greater inside exclosures and density was greater outside. Shrubs were taller inside exclosures at 3 sites grazed by sheep (Ovis aries). Perennial grasses showed no vertical height difference. Aboveground plant biomass production was different at only 1 site. Plant community diversity inside and outside exclosures were equal at 11 of 16 sites. Species richness was similar at all sites and never varied > 4 species at any site. Few changes in species composition, cover, density, and production inside and outside exclosures have occurred in 65 years, indicating that recovery rates since pre-Taylor Grazing Act conditions were similar under moderate grazing and grazing exclusion on these exclosure sites.

Verotoxin-Producing Escherichia coli in Culled Beef Cows Grazing Rangeland Forages

The objective of this study was to assess prevalence of verotoxin-producing Escherichia coli (VTEC) in culled beef cows at the time of shipping to slaughter. Feces were collected from 82 cows on eight Nevada ranches during fall and winter (from September to January) after grazing rangeland forages. A random sample (n = 154) of potential VTEC isolates were tested for verotoxicity and were screened for the presence (polymerase chain reaction) and expression (VTEC-reversed passive latex agglutination assay) of the toxin genes (i.e., VT1 and VT2). Seventeen isolates from four ranches were VTEC. Of these, four had the VT1 gene, five had the VT2 gene, seven had both genes, and one did not have either gene despite its toxicity to Vero cells. Except for one isolate (i.e., untypeable that reacted with VT1-latex beads without having VT1 gene), the genotype and phenotype data of the VTEC isolates matched. Another isolate (08:H– [nonmotile]) was verotoxic, but neither had nor expressed the toxin genes. Of the 17 isolates, four (from one cow) were O157:H7, 11 (from five cows on three ranches) were non-O157:H7 (two O8:H–, three O105:H–, three O116:H–, and three O141:H–), and two were untypeable. Because some of these VTEC serotypes (i.e., O8:H–, O141:H–, and O157:H7) are known to cause human illnesses, it is beneficial to identify VTEC-positive cows before slaughter. This is a critical step in any pre-or post-harvest strategy to minimize the risk of beef contamination with such pathogens.

Verotoxin-Producing Escherichia coli in Sheep Grazing an Irrigated Pasture or Arid Rangeland Forages

Worldwide, verotoxin-producing Escherichia coli (VTEC) have been recognized as the cause of many sporadic cases or major outbreaks of human illnesses involving consumption of contaminated meat, especially beef. Although sheep products have not been linked to reported human illnesses, their role as a food safety risk factor should not be ignored. The objective of this study was to assess VTEC prevalence in two groups of ewes (20 each) grazing an irrigated pasture or arid range in a western United States environment (Nevada) over 1 year (summer of 1999 to summer of 2000). A random sample (n = 504) of potential VTEC isolates were tested for verotoxicity and were screened for the presence (polymerase chain reaction [PCR]) and expression (VTEC-reversed passive latex agglutination assay) of the toxin genes (i.e., VT1 and VT2). Forty-one VTEC isolates (16 having only the VT1 gene and 25 having both VT1 And VT2 genes) were detected in both groups of ewes. Except for seven isolates, the genotype and phenotype data matched. All the isolates (nonmotile [H–]) were non-O157:H7 VTEC (i.e., O91:H– [n = 25], O128:H– [n = 9], and untypeable ones [n = 7]). More infected ewes (nine versus three) and different VTEC strains were found in the irrigated pasture than in the arid range. Because our ewes were shedding two VTEC serotypes known to cause human illnesses, it is beneficial to identify VTEC-positive sheep before slaughter as an initial control point before entering the food chain.

Occurrence of verotoxin-producing Escherichia coli in dairy heifers grazing an irrigated pasture

Verotoxin-producing Escherichia coli (VTEC) produce one or two toxins known as VT1 and VT2. These toxins have been associated with several human illnesses. Dairy cattle harboring VTEC represent a potential health hazard because they enter the food chain as ground beef. The objective of this study was to assess the occurrence of VTEC in dairy heifers. A total of 91 fecal samples were rectally collected during four periods (spring, summer, fall, and winter of 1999) from 23 heifers. A random sample (n=530) of potential VTEC isolates were tested for verotoxicity and were screened by the polymerase chain reaction (PCR) for presence or absence of VT1 and/or VT2 genes. Thirteen isolates from two heifers (from the winter collection) were verotoxic and were confirmed as E. coli. VTEC were only detected during winter with an occurrence rate of 9.5%. Using PCR, five isolates had the VT1 gene while the remaining eight had the VT2 gene. The sequence and expression of VT1 and VT2 genes were confirmed. No E. coli O157:H7 was detected, but serotyping revealed that the five VT1-positive isolates were O26:NM (a non-motile strain of O26). The remaining eight isolates were untypeable. Identification of VTEC-positive cattle before slaughter is a critical step in any on-farm strategy to minimize the risk of beef contamination with such pathogens.

Influence of pH treatments on survival of Escherichia coli O157:H7 in continuous cultures of rumen contents.

The pH (i.e., 5.5, 5.75, 6.0, 6.25, 6.5, 6.75, 7.0, and 7.25) effect on Escherichia coli O157:H7 in an artificial rumen model was investigated. Eight fermenters were inoculated with bovine rumen fluid and were supplied with a diet (75 g of dry matter daily in 12 equal portions [every 2 hr]) containing similar forage-to-concentrate ratio. After an adaptation period (i.e., 3 days for adjusting the rumen fluid [pH 6.2] microbial population to the test pH and 4 days for adjustment to the diet at the test pH), each fermenter was inoculated with 109 cells of E. coli O157:H7. Samples were collected hourly for 12 hr and every 2 hr for an additional 12 hr and were analyzed by flow cytometer. E. coli O157:H7 could not be quantified after 24 hr, and detection was only possible after enrichment. Because the pathogen could not be detected 5 days postinoculation (i.e., Day 13), the fermenters were reinoculated with E. coli O157:H7 on Days 17 and 22. E. coli O157:H7 numbers decreased from 106 to 104/ml of fermenter contents in a quadratic (P < 0.05) fashion over the 24-hr sampling period, and the rate of reduction was slower (P < 0.05) for pH 7.0 than for other pH treatments. Results suggested that E. coli O157:H7 population were decreased by competitive exclusion and were not affected by culture pH.

Detection of Escherichia coli O157:H7 in bovine rumen fluid and feces by flow cytometry

Abstract This study was designed to evaluate the efficacy of a flow cytometric method to detect and quantify Escherichia coli O157:H7 in the rumen fluid and feces. The rumen fluid and feces were collected from two ruminally cannulated beef steers consuming a grass-hay diet. Samples of rumen fluid and diluted (1:10) feces were filtered through cheesecloth before inoculation with E. coli O157:H7 (ATCC 43888) at various concentrations (10 3 –10 7 cells/ml). A fluorescein (FITC) conjugated anti-O157:H7 monoclonal antibody was used to label the pathogen. Negative controls were samples of rumen fluid and feces with or without a strain of non-O157:H7 E. coli (ATCC 25922). Labeled- E. coli O157:H7 were accurately enumerated at concentrations ⩾10 4 cells/ml while concentrations ⩽10 3 were only detected after enrichment. This flow cytometric method can quantify or detect E. coli O157:H7 in bovine rumen fluid and feces at concentrations commonly shed by cattle.

Introduction to the Food Safety Concerns of Verotoxin-Producing Escherichia coli

Verotoxin-producing Escherichia coli (VTEC) have emerged in the past two decades as food-borne pathogens that can cause major outbreaks of human illnesses worldwide. The number of outbreaks has increased in recent years due to changes in food production and processing systems, eating habits, microbial adaptation, and methods of VTEC transmission. The human illnesses range from mild diarrhea to hemolytic uremic syndrome (HUS) that can lead to death. The VTEC outbreaks have been attributed to O157:H7 and non-O157:H7 serotypes of E. coli. These E. coli serotypes include motile (e.g., O26:H11 and O104:H21) and nonmotile (e.g., O111:H–,0145:H–, and O157:H–) strains. In the United States, E. coli O157:H7 has been the major cause of VTEC outbreaks. Worldwide, however, non-O157:H7 VTEC (e.g., members of the 026, O103, O111, O118, O145, and O166 serogroups) have caused approximately 30% of the HUS cases in the past decade. Because large numbers of the VTEC outbreaks have been attributed to consumption of ruminant products (e.g., ground beef), cattle and sheep are considered reservoirs of these food-borne pathogens. Because of the food safety concern of VTEC, a global perspective on this problem is addressed (Exp Biol Med Vol. 228, No. 4). The first objective was to evaluate the known non-O157:H7 VTEC strains and the limitations associated with their detection and characterization. The second objective was to identify the VTEC serotypes associated with outbreaks of human illnesses and to provide critical evaluation of their virulence. The third objective was to determine the rumen effect on survival of E. coli O157:H7 as a VTEC model. The fourth objective was to explore the role of intimins in promoting attaching and effacing lesions in humans. Finally, the ability of VTEC to cause persistent infections in cattle was evaluated.

Plant Age and Growing Season Nutritional Content Relationships of Three Artemisia tridentata Subspecies

Abstract The effect of plant age on growing season chemical compositions and rumen fermentation characteristics was determined for three subspecies of big sagebrush: basin (Artemisia tridentata [Nutt.] subsp. tridentata), mountain (A. tridentata subsp. vaseyana [Rybd.] Beetle), and Wyoming (A. tridentata subsp. wyomingensis [Beetle and Young]). In vitro dry matter (IVDMD) and organic matter (IVOMD) disappearance, ammonia nitrogen (NH3N), and volatile fatty acid (VFA) content were determined at the end of two fermentation periods (24 h and 48 h) by combining rumen inocula with age-classified vegetative samples from each sagebrush subspecies. An additional one-way analysis of variance was performed to investigate potential differences among subspecies in IVDMD, IVOMD, total VFA, and NH3N following a 48-h fermentation period. Crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL) components were also compared among sagebrush subspecies. Age class responses were variable across the spectrum of sagebrush subspecies and response variables. Where plant age effects were indicated, the small numeric differences probably have little biological or ecological significance. Mountain sagebrush was lower in IVOMD and total VFA concentrations (P < 0.0001) than basin and Wyoming. NH3N concentration and CP were higher (P < 0.0001) in basin sagebrush than the other two subspecies, while Wyoming sagebrush was higher in NDF, ADF, and ADL than basin and mountain subspecies (P < 0.0001). NH3N concentration for all three subspecies was lower than the minimum level (20 mg · 100 mL−1) required for uninhibited rumen activity. Overall, this research questions the contention that older sagebrush plants offer less nutritional value than younger ones, at least for growing season conditions. The results also provide information that can be utilized in designing supplementation strategies for domestic animals on diets with characteristically high utilization of big sagebrush.

Sorbitol-Negative Shiga Toxin-Producing Escherichia coli in Beef Heifers Grazing Rangeland Forages

ers in the spring with occurrence rates of 22.7 and 8.7%, respectively. Of these isolates, 37 had and expressed Shiga Toxin 1(Stx1), 14 had and expressed Shiga Toxin 2 (Stx2), and five had but did not express stx2. Only non-O157:H7 STEC (O118:H − [non-motile] and O138:H − ) were detected. Because E. coli O118:H − is pathogenic to humans, it is important to determine its incidence in cattle and to determine the risk of beef contamination with such a pathogen.