Thermal inactivation of Yersinia enterocolitica in pork slaughter plant scald tank water

The objective of this study was to establish the time–temperature combinations required to ensure the thermal inactivation of Yersinia enterocolitica during scalding of pork carcasses. A 2 strain cocktail of Y. enterocolitica (bioserotypes 2/O:5,27 and 1A/O:6,30) was heat treated at 50, 55 and 60 °C in samples of scald tank water obtained from a commercial pork slaughter plant. Samples were removed at regular intervals and surviving cells enumerated using (i) Cefsulodin–Irgasan–Novobiocin Agar (CIN) supplemented with ampicillin and arabinose and (ii) Tryptone Soya Agar (TSA), overlaid with CIN agar with ampicillin and arabinose. The data generated was used to estimate D- and z-values and the formula D_x = log^− 1(log D₆₀ − ((t₂ − t₁)/z)) was applied to calculate thermal death time–temperature combinations from 55 to 65 °C. D₅₀, D₅₅ and D₆₀-values of 45.9, 10.6 and 2.7 min were calculated from the cell counts obtained on CIN agar, respectively. The corresponding D-values calculated from the TSA/CIN counts were 45.1, 11 and 2.5 min, respectively. The z-value was 7.8. It was concluded that a time–temperature combination of 2.7 min at 60 °C is required to achieve a 1 log reduction in Y. enterocolitica in pork scald tank water. The predicted equivalent at 65 °C was 0.6 min. This study provides data and a model to enable pork processors to identify and apply parameters to limit the risk of carcass cross-contamination with Y. enterocolitica in pork carcass scald tanks.     

Evaluation of the ISO 10273:2003 method for the isolation of human pathogenic Yersinia enterocolitica from pig carcasses and minced meat

Pig carcass swabs (n = 254) and minced meat samples (n = 82) were examined for pathogenic Yersinia enterocolitica using different routinely used enrichment protocols. All samples were obtained in the context of the official Yersinia monitoring program in Belgium. In total, 28 carcasses (11.0%) were contaminated with Y. enterocolitica bioserotype 4/O:3 and one (0.4%) with bioserotype 2/O:9. Four minced meat samples (4.9%) tested positive for Y. enterocolitica bioserotype 4/O:3. Using the ISO 10273:2003 method, eight out of the 29 Yersinia-positive carcasses (27.6%) and none of the contaminated minced meat samples (0.0%) were detected. Reducing the enrichment time in PSB from 5 to 2 days increased the number of positive samples. Overall, enrichment in PSB at 25 °C recovered more positive carcasses and minced meat samples than selective enrichment and cold enrichment. As the exclusive use of the ISO 10273:2003 method results in a strong underestimation of Y. enterocolitica positive carcasses and minced meats, efforts are needed to optimize the current version of the ISO method. In addition, isolation of pathogenic Y. enterocolitica requires experience and the use of a stereomicroscope to avoid false negative results.     

Pulsed field, PCR ribotyping and multiplex PCR analysis of Yersinia enterocolitica strains isolated from meat food in San Luis Argentina

The characterization of phenotypic and genotypic virulence markers of Yersinia enterocolitica strains belonging to biotypes (B) 1A, 2 and 3, mostly isolated from food in San Luis, Argentina, and the assessment of their genotypic diversity using PFGE and PCR ribotyping, were performed in our laboratory for the first time. Thirty five Y. enterocolitica strains, two reference strains and 33 strains isolated in our laboratory were studied. The presence of virF, ail, ystA, and myfA genes was investigated by multiplex PCR. The pathogenic potential of B1A strains, the most predominant biotype of Y. enterocolitica strains isolated from meat in our region, was investigated by simple PCR. Four B1A strains were positive for ystB gene. Four Y. enterocolitica 2/O:9 (bio/serotype) and two 3/O:5 strains isolated in our laboratory showed virulence-related results in the phenotypic tests and multiplex PCR. A good correlation between the expression of virulence markers and their corresponding genotypes was observed for most strains. Sixteen genomic types (GT) and 9 different intergenic spacer region (SR) groups were generated by PFGE and PCR ribotyping, respectively. In both cases the Y. enterocolitica 2/O:9 strains were separately clustered from 1A and 3/O:5 strains. Meat foods might be vehicles of transmission of pathogenic Y. enterocolitica strains in our region.     

Development of a multiplex and semi-nested PCR assay for detection of Yersinia enterocolitica and Aeromonas hydrophila in raw milk

Single-locus (sl), multiplex (m), and semi-nested (sn) polymerase chain reaction (PCR) procedures were developed for the detection of Yersinia enterocolitica and Aeromonas hydrophila in raw milk samples. Two oligonucleotide primers for each pathogen were used in PCR, to detect the yst gene of Y. enterocolitica and aer gene of A. hydrophila. The amplified fragment size was 145 base-pair (bp) for yst gene, and 209 bp for the aer gene. The performance of the systems were evaluated with seeded milk samples, and naturally contaminated raw milk samples. PCR results were compared with conventional cultural procedures. The limits of detection of slPCR and mPCR assays were approximately 10² cfu ml⁻¹ (0·5 cfu/PCR reaction mixture) for both pathogens in seeded raw milk. When studied with naturally contaminated raw milk samples, detection rates obtained by PCR and cultural methods were 53% and 36% for Y. enterocolitica and were 23% and 14% for A. hydrophila, respectively. These results indicate that the direct PCR analysis of raw milk can be used as a rapid and specific diagnostic method for both pathogens.     

Yersinia enterocolitica in slaughter pig tonsils: Enumeration and detection by enrichment versus direct plating culture

Tonsil samples from 139 slaughter pigs were examined for the presence of pathogenic Yersinia enterocolitica by enrichment procedures based on the standard method ISO 10273:2003. In addition, samples were tested by direct plating method to evaluate its efficiency compared to the enrichment culture methods and to quantify the level of contamination in porcine tonsils. In total, 52 samples (37.4%) were positive for pathogenic Y. enterocolitica, all belonging to bioserotype 4/O:3. Fifty out of the 52 positive samples (96.2%) were detected by direct plating. Enumeration showed an average concentration of 4.5 log₁₀ CFU g⁻¹ and 4.4 log₁₀ CFU g⁻¹ tonsil on Salmonella-Shigella-desoxycholate-calcium chloride (SSDC) and cefsulodin-irgasan-novobiocin (CIN) agar plates, respectively. The enrichment procedures recommended by the ISO 10273:2003 method were not optimal for the isolation of pathogenic Y. enterocolitica from pig tonsils: two days enrichment in irgasan-ticarcillin-potassium chlorate (ITC) broth resulted in an isolation rate of 84.6%, while 5 days enrichment in peptone-sorbitol-bile (PSB) broth recovered only 59.6% of positive samples. Reducing the enrichment time in PSB from 5 to 2 days resulted in a significantly higher recovery rate (94.2%) and might serve as an appropriate enrichment protocol for the isolation of pathogenic Y. enterocolitica from pig tonsils. Compared to enrichment culture methods, results based on direct plating can be obtained in a shorter time course and provide quantitative data that might be needed for further risk assessment studies.     

Short communication: Methicillin-resistant Staphylococcus aureus detection in US bulk tank milk

Staphylococcus aureus is a major cause of mastitis in dairy cattle. This study estimated the herd prevalence of methicillin-resistant Staph. aureus (MRSA) among US dairy herds by testing bulk tank milk (BTM) samples using genotypic and phenotypic methods. A nationally representative sample of 542 operations had BTM cultured for Staph. aureus, and 218 BTM samples were positive upon initial culture. After 4 wk to 4 mo of frozen storage, 87% of 218 samples (n = 190) were still culture positive for Staph. aureus on blood agar, but none were positive for MRSA on the selective indicator medium CHROMagar MRSA. A duplex PCR was used to detect the Staph. aureus-specific nuc gene and the methicillin resistance gene, mecA, in mixed staphylococcal isolates from the 190 BTM samples that were positive for Staph. aureus after storage. Seven samples tested positive for nuc and mecA, and 2 samples tested positive for mecA only. MecA-positive Staphylococcus spp., but not MRSA, were subsequently isolated from 5 samples, whereas neither mecA-positive Staphylococcus spp. nor MRSA was isolated from the remaining 4 samples. Presence of methicillin-resistant, coagulase-negative Staphylococcus spp. may complicate the detection of MRSA by means of PCR on BTM. Bulk tank milk in the United States is not a common source of MRSA.     

Detection of Cronobacter species in powdered infant formula by probe-magnetic separation PCR

Cronobacter species are opportunistic foodborne pathogens associated with serious infections in preterm neonates and infants. Based on the epidemiological research, infant formula products are considered to be the main source of infections from this organism. Therefore, accurate methods are required for detection of Cronobacter species. In this study, the specific probe and primers for detection of this organism were designed and verified. The probe-magnetic beads were prepared for sequence capture, followed by PCR assay to detect the target gene. This probe-magnetic separation PCR assay could detect as few as 10³ cfu/mL of Cronobacter in artificially contaminated infant formulas in less than 4 h. The combination of magnetic beads and PCR showed the potential for the detection of Cronobacter in infant formulas and may have applications in the dairy industry.     

Detection and differentiation of Vibrio spp. in seafood and fish samples with cultural and molecular methods

Vibrio spp. as natural inhabitants of sea- and brackwater of both tropical and temperate regions of the world are commonly found in different kinds of seafood. Even among the three main human pathogenic species Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus most of the isolates from seafood do not carry the different virulence factors responsible for foodborne infections. Therefore, the risk assessment of Vibrio spp. in seafood is currently based mainly on the knowledge of the genetic setting of foodborne strains. For the detection and differentiation of Vibrio spp. (V. parahaemolyticus, V. cholerae and V. vulnificus) three probe-based multiplex real-time PCR systems were developed and validated. One real-time PCR system simultaneously detects V. parahaemolyticus, V. cholerae and V. vulnificus on genus level combined with an Internal Amplification Control. The detection limit for the system was between 1 cfu/mL and 10 cfu/mL in pure culture and in different artificially contaminated sample material, e. g. prawns or Alaska Pollock. The other two PCR systems were implemented for the detection of different virulence genes of V. parahaemolyticus and V. cholerae isolates. The molecular detection systems were applied for the investigation of 338 raw and cooked seafood and fish samples for the presence of the different Vibrio spp. The collected data indicate that the PCR systems can be useful for rapid detection and differentiation of Vibrio spp. in different food matrices as basis for a preventive consumer protection policy.     

Probiotic beverage from cashew apple juice fermented with Lactobacillus casei

This study optimized the conditions of Lactobacillus casei NRRL B-442 cultivation in cashew apple juice, as well as, determined the proper inoculum amount and fermentation time. Moreover, it was investigated the survivability ability of L. casei in cashew apple juice during refrigerated storage (4 °C) for 42 days. The optimum conditions for probiotic cashew apple juice production were initial pH 6.4, fermentation temperature of 30 °C, inoculation level of 7.48 Log CFU/mL (L. casei) and 16 h of fermentation process. It was observed that the L. casei grew during the refrigerated storage. Viable cell counts were higher than 8.00 Log CFU/mL throughout the storage period (42 days). The values of lightness, yellowness and total color change increased and the values of redness reduced along the fermentation and refrigerated storage periods. The fermented juice with L. casei is a good and healthy alternative functional food containing probiotics. Cashew apple juice showed to be as efficient as dairy products for L. casei growth.     

Comparative efficacy of Zataria multiflora Boiss., Origanum compactum and Eugenia caryophyllus essential oils against E. coli O157:H7, feline calicivirus and endogenous microbiota in commercial baby-leaf salads

Ready-to-eat salads using baby-leaf and multi-leaf mixes are one of the most promising developments in the fresh-cut food industry. There is great interest in developing novel decontamination treatments, which are both safe for consumers and more efficient against foodborne pathogens. In this study, emulsions of essential oils (EOs) from Origanum compactum (oregano), Eugenia caryophyllus (clove), and Zataria multiflora Boiss (zataria) were applied by spray (0.8 ml) after the sanitizing washing step. The aim was to investigate their ability to control the growth of potentially cross-contaminating pathogens and endogenous microbiota in commercial baby leaves, processed in a fresh-cut produce company. Zataria EO emulsions of 3%, 5% and 10% reduced Escherichia coli O157:H7 by 1.7, 2.2 and 3.5 log cfu/g in baby-leaf salads after 5 days of storage at 7 °C. By contrast, reductions in E. coli O157:H7 counts remained the same when clove was applied at concentrations of 5% and 10% (2.5 log cfu/g reduction). Oregano (10%) reduced inoculated E. coli O157:H7 counts in baby-leaf salads by a maximum of 0.5 log cfu/g after 5 days of storage. Zataria showed strong antimicrobial efficacy against E. coli O157:H7 and also against the endogenous microbiota of baby-leaf salads stored for 9 days. Feline calicivirus (FCV), a norovirus surrogate, survived on inoculated baby-leaf salads during refrigerated storage (9 days at 7 °C) regardless of treatment. Refrigeration temperatures completely annulled the effectiveness of the EOs against FCV inoculated in baby-leaf salads as occurred in FCV cultures. This study shows that EOs, and zataria in particular, have great potential use as an additional barrier to reduce contamination-related risks in baby-leaf salads. However, further research should be done into foodborne viruses in order to improve food safety.     

Detection of Pathogenic Yersinia enterocolitica in Meat using Real-Time PCR

Yersinia enterocolitica is an important zoonosis, which can cause disease in humans and animals. The culture methods available for detection of Y. enterocolitica in food samples are time-consuming and seldom successful. Using DNA-based methods, like PCR, this pathogen can be detected more rapidly and with greater sensitivity. The aim of this study was to establish a rapid and accurate real-time PCR method to detect pathogenic Y. enterocolitica in pork. The chromosomal ail−gene, which is only present in pathogenic Y. enterocolitica strains, was used as DNA target for the 5' nuclease PCR assay. The probe was labelled at the 5' end with the fluorescent reporter dye (FAM) and at the 3' end with the quencher dye (TAMRA). A 2-step protocol with 45 cycles was used in a multicolour real-time PCR detection system. A Ct value over 40 indicated a negative result. The DNA extraction procedure for the natural samples was rapid and simply. This qualitative real-time PCR method was shown to be specific and sensitive. Detection rate of ail-positive Y. enterocolitica in 200 pig tonsils was 88 % and 35 % with PCR and culture methods, respectively. When 100 raw pork samples were studied, 7 were positive with PCR and all were culture negative.     

Short communication: An evaluation of the effectiveness of Lactobacillus buchneri 40788 to alter fermentation and improve the aerobic stability of corn silage in farm silos

The objective of this study was to determine if the effects of inoculation with Lactobacillus buchneri 40788 were detectable when applied to whole-plant corn stored in farm silos. Corn silage was randomly sampled from farms in Wisconsin, Minnesota, and Pennsylvania, and was untreated (n = 15) or treated with an inoculant (n = 16) containing L. buchneri 40788 alone or this organism combined with Pediococcus pentosaceus during May and June 2007. Corn silage that was removed from the silo face during the morning feeding was sampled, vacuum-packed, and heat sealed in polyethylene bags and shipped immediately to the University of Delaware for analyses. Silage samples were analyzed for dry matter (DM), nutrient composition, fermentation end-products, aerobic stability, and microbial populations. The population of L. buchneri in silages was determined using a real-time quantitative PCR method. Aerobic stability was measured as the time after exposure to air that it took for a 2°C increase above an ambient temperature. The DM and concentrations of lactic and acetic acids were 35.6 and 34.5, 4.17 and 4.85, and 2.24 and 2.41%, respectively, for untreated and inoculated silages and were not different between treatments. The concentration of 1,2-propanediol was greater in inoculated silages (1.26 vs. 0.29%). Numbers of lactic acid bacteria determined on selective agar were not different between treatments. However, the numbers of L. buchneri based on measurements using real-time quantitative PCR analysis were greater and averaged 6.46 log cfu-equivalents/g compared with 4.89 log cfu-equivalent for inoculated silages. There were fewer yeasts and aerobic stability was greater in inoculated silages (4.75 log cfu/g and 74 h of stability) than in untreated silages (5.55 log cfu/g and 46 h of stability). This study supports the effectiveness of L. buchneri 40788 on dairy farms.     

A filtration-based real-time PCR method for the quantitative detection of viable Salmonella enterica and Listeria monocytogenes in food samples

We developed a novel filtration-based method that can eliminate dead or severally damaged Salmonella enterica and Listeria monocytogenes in food samples. This new method can recover all viable bacteria in less than 30 min, and can be coupled with a subsequent bacterial DNA extraction and real-time PCR. No statically significant differences (p < 0.01) were found between real-time PCR results obtained separately from S. enterica and L. monocytogenes when different ratios of living and dead cells were used. The analytical sensitivity in both cases was 1 genome equivalent (GE), and the quantification was linear (R² > 0.9969) over a 5-log dynamic range with PCR efficiencies >0.9754. When compared with the standard microbiological methods for the detection of these foodborne pathogens, the relative accuracy was excellent ranging from 95.72% to 104.48%. Finally, we applied the pre-treatment method to the direct detection of viable forms of these foodborne pathogens in food samples using yogurt as a model, the results being similar to those obtained using pure cultures.     

Spray application of liquid smoke to reduce or eliminate Listeria monocytogenes surface inoculated on frankfurters

In a simulated post process contamination scenario liquid smoke was sprayed on the frankfurters after peeling, and then inoculated with Listeria monocytogenes (Lm). Samples that did not receive a liquid smoke spray remained at approximately 2 log cfu/cm² during the 48 h of storage while the levels on the liquid smoke treated frankfurters continued to decline until they were below detection level (1 cfu/100 cm²). A shelf-life study lasting 140 days indicated that liquid smoke suppressed the growth of Lm for up to 130 days. An application of 2 or 3 ml liquid smoke at packaging resulted in at least a 1 log reduction of Lm within 12 h post packaging.     

A novel real-time polymerase chain reaction-based method for the detection and quantification of lactose-fermenting Enterobacteriaceae in the dairy and other food industries

The presence of lactose-fermenting Enterobacteriaceae and coliforms is routinely assessed to determine the hygienic quality of water and foods, particularly dairy products. This paper reports the use of lacZ-specific primers in an SYBR green I-based real-time PCR method for the easy and rapid detection of coliforms in dairy products. A large number of bacterial species were assayed to establish the specificity of the method. The sensitivity of the method was assessed using artificially contaminated cheeses. The limit of detection was 1 coliform cell in cheese samples enriched for 8 h in a culture medium. The entire procedure, including sample processing, enrichment, DNA extraction, and real-time PCR amplification, can be completed within 10 to 12 h, making it a single-day assay.     

Antibacterial effects of natural tenderizing enzymes on different strains of Escherichia coli O157:H7 and Listeria monocytogenes on beef

This study determined the efficacy of actinidin and papain on reducing Listeria monocytogenes and three mixed strains of Escherichia coli O157:H7 populations on beef. The average reduction of E. coli O157:H7 was greater than that of L. monocytogenes and higher concentrations of either protease yielded greater reduction in bacterial populations. For instance, actinidin at 700 mg/ml significantly (p ≤ 0.05) reduced the population of L. monocytogenes by 1.49 log cfu/ml meat rinse after 3 h at 25 & 35 °C, and by 1.45 log cfu/ml rinse after 24 h at 5 °C, while the same actinidin concentration significantly reduced the populations of three mixed strains of E. coli O157:H7 by 1.81 log cfu/ml rinse after 3 h at 25 & 35 °C, and 1.94 log cfu/ml rinse after 24 h at 5 °C. These findings suggest that, in addition to improving the sensory attributes of beef, proteolytic enzymes can enhance meat safety when stored at suitable temperatures.     

Bifidobacterium Bb-12 microencapsulated by spray drying with whey: Survival under simulated gastrointestinal conditions,tolerance to NaCl,and viability during storage

Bifidobacterium Bb-12 was microencapsulated by spray drying with whey. This present work investigated the survival of these microcapsules under simulated gastrointestinal conditions, as well as their tolerance to NaCl and their viability during storage. The results showed a small decrease in the viability of microencapsulated Bifidobacterium at low pH. In relation to the exposure of Bifidobacterium to bile, microencapsulation with whey did not protect the probiotic cells; however, the viability of the microcapsules remained >6 log cfu/g, even after 24 h of incubation at the highest bile concentration analyzed. No growth was noted with either the free cells or the microencapsulated cells on MRS-LP with NaCl. The viability of the microcapsules stored at 4 °C remained high and constant for 12 weeks. When the microcapsules were added to a dairy dessert, the probiotic count remained above 7 log cfu/g for 6 weeks. Therefore, whey is a promising encapsulating agent for Bifidobacterium Bb-12.     

Technical note: Development of a challenge model for Streptococcus uberis mastitis in dairy heifers

A challenge model for experimentally inducing Streptococcus uberis mastitis in bred dairy heifers was developed. Qualifying heifers (n = 7) exhibited antibody titers of < 1:10,000 against Strep. uberis antigens and were free of intramammary infections (IMI). Two contralateral quarters of each heifer were assigned to receive an infusion of Strep. uberis (1,000 to 2,000 cfu); remaining quarters served as unchallenged controls. For a successful challenge and infection, 3 of 4 consecutive mammary secretion samples had to culture positive for Strep. uberis. Six of the 7 heifers were challenged successfully in both infused quarters with a mean dose of 1,080 cfu; once confirmed, infections were treated with a one-time infusion of nonlactating cow therapy. Before challenge, mammary secretion leukocyte counts averaged 8.4 × 10⁶/mL in all quarters. At 24 h after challenge, leukocyte count increased to 18.4 × 10⁶/mL in challenged quarters, peaking on d 5 at 24.3 × 10⁶/mL; unchallenged quarters remained at ≤ 10.4 × 10⁶/mL, but increased to 15.2 × 10⁶/mL on d 7 and then decreased. Before challenge, macrophages predominated (81%) in mammary secretions followed by lymphocytes (15.3%) and neutrophils (3.7%). By 24 h after challenge, neutrophils increased in challenged quarters and predominated for the duration of the trial (65.3 to 70%), whereas macrophages predominated in unchallenged control quarters (65.2 to 75.2%). The challenge model was successful in establishing Strep. uberis IMI in 85.7% of animals, and IMI were controlled (100% cure) by administering nonlactating cow therapy. All heifers calved free of IMI and antimicrobial residues, with milk production similar to that of herd mates and with somatic cell counts (SCC) < 200,000 cells/mL.     

Prevalence and comparison of Streptococcus infantarius subsp. infantarius and Streptococcus gallolyticus subsp. macedonicus in raw and fermented dairy products from East and West Africa

Streptococcus infantarius subsp. infantarius (Sii) and Streptococcus gallolyticus subsp. macedonicus are members of the Streptococcus bovis/Streptococcus equinus complex (SBSEC) associated with human infections. SBSEC-related endocarditis was furthermore associated with rural residency in Southern Europe. SBSEC members are increasingly isolated as predominant species from fermented dairy products in Europe, Asia and Africa. African variants of Sii displayed dairy adaptations to lactose metabolism paralleling those of Streptococcus thermophilus including genome decay. In this study, the aim was to assess the prevalence of Sii and possibly other SBSEC members in dairy products of East and West Africa in order to identify their habitat, estimate their importance in dairy fermentation processes and determine geographic areas affected by this potential health risk. Presumptive SBSEC members were isolated on semi-selective M17 and SM agar media. Subsequent genotypic identification of isolates was based on rep-PCR fingerprinting and SBSEC-specific16S rRNA gene PCR assay. Detailed identification was achieved through application of novel primers enhancing the binding stringency in partial groES/groEL gene amplification and subsequent DNA sequencing. The presence of S. thermophilus-like lacS and lacZ genes in the SBSEC isolates was determined to elucidate the prevalence of this dairy adaptation. Isolates (n = 754) were obtained from 72 raw and 95 fermented milk samples from Côte d'Ivoire and Kenya on semi-selective agar media. Colonies of Sii were not detected from raw milk despite high microbial titers of approximately 10⁶ CFU/mL on M17 agar medium. However, after spontaneous milk fermentation Sii was genotypically identified in 94.1% of Kenyan samples and 60.8% of Kenyan isolates. Sii prevalence in Côte d'Ivoire displayed seasonal variations in samples from 32.3% (June) to 40.0% (Dec/Jan) and isolates from 20.5% (June) to 27.7% (Dec/Jan) present at titers of 10⁶–10⁸ CFU/mL. lacS and lacZ genes were detected in all Kenyan and 25.8% (June) to 65.4% (Dec/Jan) of Ivorian Sii isolates. Regional differences in prevalence of Sii and dairy adaptations were observed, but no clear effect of dairy animal, fermentation procedure and climate was revealed. Conclusively, the high prevalence of Sii in Kenya, Côte d'Ivoire in addition to Somalia, Sudan and Mali strongly indicates a pivotal role of Sii in traditional African dairy fermentations potentially paralleling that of typical western dairy species S. thermophilus. Putative health risks associated with the consumption of high amounts of live Sii and potential different degrees of evolutionary adaptation or ecological colonization require further epidemiologic and genomic investigations, particularly in Africa.     

Feeding saponin-containing Yucca schidigera and Quillaja saponaria to decrease enteric methane production in dairy cows

An experiment was conducted in vitro to determine whether the addition of saponin-containing Yucca schidigera or Quillaja saponaria reduces methane production without impairing ruminal fermentation or fiber digestion. A slightly lower dose of saponin was then fed to lactating dairy cows to evaluate effects on ruminal fermentation, methane production, total-tract nutrient digestibility, and milk production and composition. A 24-h batch culture in vitro incubation was conducted in a completely randomized design with a control (no additive, CON) and 3 doses of either saponin source [15, 30, and 45 g/kg of substrate dry matter (DM)] using buffered ruminal fluid from 3 dairy cows. The in vivo study was conducted as a crossover design with 2 groups of cows, 3 treatments, and three 28-d periods. Six ruminally cannulated cows were used in group 1 and 6 intact cows in group 2 (627 ± 55 kg of body weight and 155 ± 28 d in milk). The treatments were 1) early lactation total mixed ration, no additive (control; CON); 2) CON diet supplemented with whole-plant Y. schidigera powder at 10 g/kg of DM (YS); and 3) CON diet supplemented with whole-plant Q. saponaria powder at 10 g/kg of DM (QS). Methane production was measured in environmental chambers and with the sulfur hexafluoride (SF₆) tracer technique. In vitro, increasing levels of both saponin sources decreased methane concentration in the headspace and increased the proportion of propionate in the buffered rumen fluid. Concentration of ammonia-N, acetate proportion, and the acetate:propionate ratio in the buffered rumen fluid as well as 24-h digestible neutral detergent fiber were reduced compared with the CON treatment. Medium and high saponin levels decreased DM digestibility compared with the CON treatment. A lower feeding rate of both saponin sources (10 g/kg of DM) was used in vivo in an attempt to avoid potentially negative effects of higher saponin levels on feed digestibility. Feeding saponin did not affect milk production, total-tract nutrient digestibility, rumen fermentation, or methane production. However, DM intake was greater for cows fed YS and QS than for CON cows, with a tendency for greater DM intake for cows fed YS compared with those fed QS. Consequently, efficiency of milk production (kg of milk/kg of DM intake) was lower for cows fed saponin compared with controls. The results show that although saponin from Y. schidigera and Q. saponaria lowered methane production in vitro, the reduction was largely due to reduced ruminal fermentation and feed digestion. Feeding a lower dose of saponin to lactating dairy cows avoided potentially negative effects on ruminal fermentation and feed digestion, but methane production was not reduced. Lower efficiency of milk production of cows fed saponin, and potential reductions in feed digestion at high supplementation rates may make saponin supplements an unattractive option for lowering methane production in vivo.