Short communication: Effect of pomegranate-residue supplement on Cryptosporidium parvum oocyst shedding in neonatal calves

Cryptosporidium parvum is considered one of the most common enteropathogens, responsible for the high incidence of diarrhea and deleterious implications on immunity and health in neonatal calves. The pomegranate is well known for its health-promoting properties. Two experiments were designed to test the antiparasitical and antidiarrheal effects of concentrated pomegranate extract (CPE) supplement in milk in neonatal Holstein calves. Forty-one calves were randomly divided into control (n = 20) and treatment (n = 21) groups. For the first experiment, the treatment group was supplemented with 3.75% CPE in the daily milk ration, between 3 and 14 d of age, whereas the control group received only milk. Fecal samples were collected between d 5 and 13 to quantify Cryptosporidium oocysts, and the duration and intensity of diarrhea were evaluated. Reduced fecal oocyst count and diarrhea intensity and duration were revealed in the 3.75% CPE calves. No difference was noted in average daily gain between groups. In a second experiment, which was designed to test the effect of a lower CPE concentration (0.6% of daily milk allocation), no effects on fecal oocyst count and average daily gain were observed. However, compared with control, the lower CPE group was characterized by a shorter duration of diarrhea and higher weight gain among males at 14 d of age. These results suggest that the CPE supplement-to-calf milk ratio may potentially alleviate intestinal morbidity caused by Cryptosporidium.     

Effect of halofuginone lactate on the occurrence of Cryptosporidium parvum and growth of neonatal dairy calves

Thirty-one Holstein bull calves were purchased at birth from 3 dairy farms in Eastern Ontario. Each calf was assigned at random to oral treatment with either 5 mg of halofuginone lactate in 10.0 mL of aqueous carrier solution (Halocur, base comprised 10 mg of benzoic acid, 100 mg of lactic acid, and 0.3 mg of tartrazine) or 10 mL of placebo (Halocur base minus the active ingredient, halofuginone lactate) administered 15 to 30 min after morning milk feeding for the first 7 d of life. Intakes of milk, calf starter, and water, and fecal consistency score were recorded daily for 56 d. Calf weights were recorded weekly for 56 d. Fecal samples were taken from all calves at approximately 2, 7, 14, 21, and 28 d of age for isolation of Cryptosporidium parvum oocysts. Logistic and linear regression analyses were used to assess the effect of treatment on the incidence of diarrhea and C. parvum infection status. The odds of C. parvum shedding among calves in the halofuginone lactate-treated group was 70% lower than the odds of shedding among calves in the placebo group. In calves treated with halofuginone lactate, no oocyst shedding occurred until 2 wk of age, whereas 12.5% of calves in the placebo group began shedding oocysts during wk 1. From all ages of placebo-treated calves, 31 of 73 samples (42.5%) were positive for C. parvum, whereas only 15 of 67 samples (22.4%) from all ages of halofuginone lactate-treated calves tested positive. The largest number of C. parvum-positive samples occurred in the third week of life. There was a significant delay of 3.1 d in the incidence of diarrhea among calves treated with halofuginone lactate. Intake of milk and starter, body weight gains, and age at weaning were not significantly different between treatment groups.     

Feeding activated charcoal from bark containing wood vinegar liquid (nekka-rich) is effective as treatment for cryptosporidiosis in calves

The effect of activated charcoal containing wood vinegar liquid (Nekka-Rich) on Cryptosporidium parvum was evaluated in vitro and in vivo. First, the adsorption of C. parvum by the activated charcoal component of Nekka-Rich was examined. When mixed, C. parvum oocysts were effectively adsorbed by activated charcoal. Next, the effect of the wood vinegar liquid component of Nekka-Rich was assessed. Wood vinegar liquid had an antiprotozoan activity against C. parvum oocysts. Finally, the anticryptosporidial effect of Nekka-Rich was evaluated in calves experimentally infected with C. parvum. Six neonatal calves at 7 d of age were orally infected with 1 × 10⁵ oocysts of C. parvum. When diarrhea was observed (on d 3 after the experimental infection), 3 calves received a milk replacer containing 10 g of Nekka-Rich at 8-h intervals for 4 consecutive days, and 3 control calves were given a milk replacer without Nekka-Rich at 8-h intervals for 4 consecutive days. Calves fed milk with Nekka-Rich showed recovery from diarrhea 1 d after the start of treatment. Significantly less fecal excretion of C. parvum oocysts was observed 1 d after treatment in calves fed Nekka-Rich. On d 2 from the beginning of treatment with Nekka-Rich (on d 5 after the experimental infection), oocysts were not isolated from fecal samples. On the other hand, nontreated control calves developed severe diarrhea, and fecal excretion of oocysts was consistently observed. Calves in both groups were killed for collection of samples, which were subjected to scanning electron microscopy. Scanning electron microscopy revealed adherence of C. parvum oocysts to the intestinal epithelial surface of nontreated control calves, whereas a lack of C. parvum adhesion was observed in calves fed Nekka-Rich. These results suggest that Nekka-Rich would be useful for the treatment of cryptosporidiosis in neonatal calves.     

Effect of nitazoxanide on cryptosporidiosis in experimentally infected neonatal dairy calves

Cryptosporidium is a zoonotic protozoan that is most often diagnosed in association with diarrhea in 1- to 3-wk-old dairy calves. There are neither consistently effective nor approved antimicrobial drugs for treatment in animals. The objective of this study was to test nitazoxanide (NTZ) as a treatment for cryptosporidiosis in experimentally infected dairy calves. A randomized, controlled, and blinded trial was performed using Holstein bull calves obtained from a large commercial dairy. All births were attended by study personnel and calves were fed 4 L of heat-treated colostrum within 1 h of birth. Calves were randomly assigned to treatment or placebo group and maintained for a 32-feeding (16 d) study period. Twenty-three calves were enrolled with 3 lost to follow up. Thirteen calves were assigned to the treatment group and 7 calves to the placebo group. All calves were inoculated with 1 × 10⁶ viable Cryptosporidium parvum oocysts at feeding 3. Treatment was a commercially available NTZ product and the placebo was the carrier of the same product. Nitazoxanide was administered at 1.5 g twice per day for 5 d. Nitazoxanide or placebo treatment began after feeding 10 and when the fecal score was greater than 1 out of 3. Outcome measurements included twice-daily fecal and health scores and a once-daily oocyst count by an immunofluorescent antibody assay. Data were analyzed by nonparametric and time-to-event methods. Measures of passive transfer of antibodies, initial body weight, and onset of oocyst shedding were not different between treatment and control calves. Eighty-five percent of the NTZ-treated calves stopped shedding oocysts by the end of the observation period whereas only 15% of the placebo group stopped shedding. The median number of feedings with a fecal score equal to 3 was 2 in the NTZ group while it was 6 in the placebo group. Calves receiving NTZ were 0.13 times as likely to have severe and sustained diarrhea than control calves (95% confidence interval, 0.02-0.98). Treating calves with NTZ reduced the duration of oocyst shedding and improved fecal consistency.     

Effects of mannan-oligosaccharide and Bacillus subtilis supplementation to preweaning Holstein dairy heifers on body weight gain,diarrhea, and shedding of fecal pathogens

The objective of this clinical trial was to evaluate the effectiveness of probiotic, prebiotic, and synbiotic supplementation on average daily weight gain (ADG), duration of diarrhea, age at incidence of diarrhea, fecal shedding of Cryptosporidium oocysts, enteric pathogens, and the odds of pneumonia in preweaning dairy heifer calves on a commercial dairy. Feeding prebiotics and probiotics may improve health and production of calves. Hence, healthy Holstein heifer calves (n = 1,801) from a large California dairy were enrolled at 4 to 12 h of age and remained in this study until weaning at 60 d of age. Calves were block randomized to 1 of 4 treatments: (1) control, (2) yeast culture enriched with mannan-oligosaccharide (prebiotic), (3) Bacillus subtilis (probiotic), and (4) combination of both products (synbiotic), which were fed in milk twice daily from enrollment until weaning. Serum total protein at enrollment and body weight at 7, 42, and 56 d of age were measured. Fecal consistency was assessed daily for the entire preweaning period. A subgroup of 200 calves had fecal samples collected at 7, 14, 21, and 42 d for microbial culture and enumeration of Cryptosporidium oocysts by direct fluorescent antibody staining. Synbiotic-treated calves had 19 g increased ADG compared with control calves for overall ADG, from 7 to 56 d. From 42 to 56 d, prebiotic-treated calves had 85 g greater ADG and synbiotic-treated calves had 78 g greater ADG than control calves. There was no difference in duration of the first diarrhea episode, hazard of diarrhea, or odds of pneumonia per calf with treatment. Probiotic-treated calves had 100 times lower fecal shedding of Cryptosporidium oocysts at 14 d and prebiotic-treated calves had fewer Escherichia coli and pathogenic E. coli at 42 d compared with control calves. Although there were no effects on duration of diarrhea or pneumonia incidence, greater ADG in the late preweaning period may reflect treatment effects on enteric pathogens during the rearing process. The decreased shedding of Cryptosporidium should reduce infectious pressure, environmental contamination, and public health risks from Cryptosporidium. Our findings suggest ADG and potential health benefits for calves fed prebiotics, probiotics, and synbiotics and can help the dairy industry make informed decisions on the use of these products in dairy production.     

Microbiological Aspects of Water Quality and Health

Historically, typhoid and cholera are well-recognized waterborne infections. Advances in laboratory techniques over the past 20 years have enabled a wide range of other faccally-excreted bacteria, viruses and protozoa to be identified as causing gastrointestinal infections. Record outbreaks are referred to where some of these micro-organisms have been spread by the contamination of drinking water, either at source or during distribution. This has usually happened when the water was untreated, or where there have been failures of the disinfection process or breaches in the integrity of the distribution system. Recent waterborne outbreaks of cryptosporidiosis have underlined the importance of adequate filtration in the prevention of waterborne transmission of this protozoon, the oocysts of which are completely resistant to the levels of chlorine that can be attained in water treatment practice. An outline is given of the current processes by which pathogenic micro-organisms are excluded from drinking-water supplies; their potential deficiencies for dealing with some of the 'newer' waterborne pathogens are also discussed.     

隐孢子虫介水传播与在水环境中存在水平

介绍了隐孢子虫的生活形态、致病症状和介水传播特点,及卵囊在水环境中存在水平和对饮用水安全的威胁;提出了以饮用水为媒介导致隐孢子虫病暴发流行的控制屏障碍环节和防治方法.隐孢子虫是一种致病的肠道寄生原虫,可引起腹泻,目前尚无特效疗法,对于免疫功能缺陷者会危及生命.以水为媒介是隐孢子虫病感染人体的重要传播途径.英美等国隐孢子虫病暴发流行的事件表明,以饮用水为媒介造成隐孢子虫病暴发流行的危险性最大.调查结果表明,水环境中存在隐孢子虫卵囊传染源的可能性较大,存在水平较高;在饮用水供水系统中也检测出隐孢子虫卵囊.隐孢子虫卵囊抗灭活能力极强,氯等消毒剂和一般剂量不能将其杀死.传统的饮用水处理工艺及消毒方法不能有效地祛除隐孢子虫.根据我国目前饮用水处理工艺技术水平,及水源污染和供水水质现状,存在介水隐孢子虫病暴发流行的隐患极大.有必要采取多屏障控制措施,特别是采取臭氧和紫外线等可靠灭活工艺,防止介水隐孢子虫病暴发流行.