In vitro activity of 24 antimicrobial agents against Staphylococcus and Streptococcus isolated from diseased animals in Japan

A total of 88 Staphylococcus and 61 Streptococcus isolates from diseased animals throughout Japan were examined in 2000 for the minimum inhibitory concentrations of 24 different antimicrobials by the agar dilution method standardized by the Japanese Society of Chemotherapy. The resistance rates to aminobenzylpenicillin (36.4%) and benzylpenicillin (35.2%) were high in Staphylococcus isolates, and those to oxytetracycline (45.9%) and kanamycin (21.3%) were high in Streptococcus isolates. Two isolates resistant to oxacillin harbored the mecA gene. One was Staphylococcus epidermidis derived from a pig with arthritis, and the other Staphylococcus cohnii from a head of cattle with mastitis.     

Antimicrobial susceptibility of Mannheimia haemolytica isolates from cattle in Japan from 2001 to 2002

A total of 27 clinical isolates of Mannheimia haemolytica from cattle in Japan from 2001 to 2002 were examined for antimicrobial susceptibility to 25 antimicrobial agents. The minimum inhibitory concentrations of 25 different antimicrobials were determined by an agar dilution method according to the guidelines of the National Committee for Clinical Laboratory Standards. Of the 27 isolates, seven isolates (26.9%) were resistant to at least one of the 25 drugs and resistance rates ranged from 3.7 to 18.5%. Resistance rates to dihydrostreptomycin (18.5%), oxytetracycline (11.1%), and doxycycline (11.1%) were relatively high and those to the remaining drugs were less than 10%.     

Microscopic structure of the large intestinal mucosa in piglets during an antibiotic-associated diarrhea

Antibiotic-associated diarrhea (AAD) is caused by the treatments of broad-spectrum antimicrobials that seriously affect the activity and composition of the large intestinal microflora. The pathogenic bacteria or low concentration of short-chain fatty acids (SCFAs) has been repeatedly discussed in relation to AAD. Recently, we reported the detection of a large amount of succinate and lactate in the diarrheal feces in AAD-induced piglets. In this study, we investigated histologically the large intestinal mucosa in AAD-induced piglets, in which succinate and lactate were accumulated. AAD was induced in the piglets by an oral dose of polymyxin B sulfate (PL) or by an intra-muscular injection of enrofloxacin (ERFX). When the piglets were defecating diarrheal feces with a high concentration of succinate and/or lactate, the large intestine was removed and separated into four segments (cecum, gyri centripetales, gyri centrifugales, and rectum). Healthy piglets were used as the control. In the AAD-induced piglets, the lamina propria was edematous in the gyri centripetales. Piglets treated with ERFX were also edematous in gyri centrifugales. These edematous lamina propria contained larger amounts of inflammatory cells than observed in control tissues. ERFX-treated piglets had a more shallow crypt than PL-treated and control piglets. The mucosal tissue of the large intestine was more seriously damaged in the ERFX- than in the PL-treated piglets, which might have been caused by the high succinate and low SCFAs concentration in the digesta.     

A cell wall preparation of Enterococcus faecalis strain EC-12 stimulates β-defensin expression in newly hatched broiler chicks

In our previous study, a cell wall preparation of Enterococcus faecalis strain EC‐12 (EC‐12) prevented the colonization of vancomycin‐resistant enterococci (VRE) in newly hatched broiler chicks. Relatively early prevention against VRE colonization by EC‐12 administration may be related to the enhanced innate immune system. In this study we examined the effect of EC‐12 on the kinetics of immunoglobulin A (IgA) concentration in the cecal digesta of chicks from 1 day to 14 days old to evaluate humoral immunity. We also examined the effect of EC‐12 on β‐defensin mRNA expression and lysozyme production. The IgA concentration in the cecal digesta showed a decline in chicks from 2 to 8 days old and then a recovery up to when they were 14 days old. The concentration tended to be higher in EC‐12‐administered chicks than in control chicks at 14 days old. EC‐12 administration did not stimulate the lysozyme activities in the alimentary tract. However, β‐defensin mRNA expression in the tongue and the bursa of Fabricius was higher in chicks administered EC‐12 than in the control chicks at 5 days old. These results indicate that β‐defensin may be one of the major defense mechanisms inhibiting VRE colonization in young chicks.     

The preventive effect of Bacillus subtilus strain DB9011 against experimental infection with enterotoxcemic Escherichia coli in weaning piglets

Porcine edema disease (ED) is caused by Shiga toxin 2e‐producing Escherichia coli (STEC). Post‐weaned piglets often suffer from ED as a result of intestinal infection with STEC, which causes impaired growth performance and high mortality. Antimicrobial therapy is a curative treatment for piglets infected with STEC, but the emergence of antimicrobial‐resistant STEC has become a serious problem for Japanese pig farmers. Therefore, an alternative strategy other than antimicrobial therapy is needed for the prevention or treatment of ED. In this study, we evaluated the effect of oral administration of Bacillus subtilis DB9011 (DB9011) to prevent the experimental infection of STEC in weaning piglets. Eight 21‐day‐old piglets were divided into two groups: STEC challenge with the basal diet, and STEC challenge with DB9011 supplemented diet. The challenge was carried out when the animals were 25, 26 and 27 days old using STEC contained in capsules resistant against gastric digestion. All pigs were euthanized at 36 days of age. DB9011 improved the symptoms of ED and decreased the number of STEC in the ileal digesta and feces. Accordingly, oral administration of DB9011 in weaned piglets prevents ED through the suppression of the growth of STEC in the ileum.     

Shotgun proteomic analysis of porcine colostrum and mature milk

The epitheliochorial nature of the porcine placenta prevents the transfer of maternal immunity. Therefore, ingestion of the colostrum immediately after birth is crucial for neonatal piglets to acquire passive immunity from the sow. We performed a shotgun proteomic analysis of porcine milk to reveal in detail the protein composition of porcine milk. On the basis of the Swiss‐Prot database, 113 and 118 proteins were identified in the porcine colostrum and mature milk, respectively, and 50 of these proteins were common to both samples. Some immune‐related proteins, including interleukin‐18 (IL‐18), were unique to the colostrum. The IL‐18 concentration in the colostrum and mature milk of four sows was measured to validate the proteomic analysis, and IL‐18 was only detected in the colostrum (191.0 ± 53.9 pg/mL) and not in mature milk. In addition, some proteins involved in primary defense, such as azurocidin, which has never been detected in any other mammal's milk, were also identified in the colostrum.     

Oral administration of Lactobacillus plantarum Lq80 and Megasphaera elsdenii iNP‐001 induces efficient recovery from mucosal atrophy in the small and the large intestines of weaning piglets

Weaning causes atrophy of intestinal mucosa and a drop of IgA protection in piglets which increases vulnerability to pathogenic infections. Probiotic lactobacilli may support recovery from such weaning stresses. Butyrate‐produce bacteria may support the growth of colonic mucosa. Megasphaera elsdenii, a lactate‐utilizing butyrate producer, may help butyrate production particularly when combined with lactobacilli. Weaned piglets (Experiment 1: 20 days old, Experiment 2: 28 days old) were orally dosed once a day with either (L) 10¹⁰ (cell/dose) L. plantarum Lq80, or (LM) 10¹⁰ (cell/dose) Lq80 with 10⁹ (cell/dose) M. elsdenii iNP‐001. Lq80 was contained in capsules resistant to gastric digestion. M. elsdenii was contained in capsules resistant to gastric and intestinal digestion. An untreated control (C) was also prepared. After 2 weeks of administration, L. plantarum enhanced the recovery from the villous atrophy in both experiments. The rectal and colonic IgA tended to be higher in L and LM than in C in Experiment 1. Colonic butyrate was higher in LM than in the others in Experiment 1. The thickness of the colonic mucosa was greater in LM than in the others in Experiment 1. In early weaned piglets, the effects of L. plantarum and M. elsdenii were clear.     

Effects of replacing fish oil with canola oil on the growth performance,fatty acid composition and metabolic enzyme activity of juvenile yellowtail Seriola quinqueradiata (Temminck & Schlegel, 1845)

In this study, fish oil (FO) was replaced with canola oil (CO) in juvenile yellowtail diets to establish the optimal replacement levels that ensure adequate feed quality for aquaculture. Juvenile yellowtails (initial body weight: 104.7 g) were fed one of four diets for 10 weeks: 100% FO (FO group) or FO replacement with 25%, 50% or 100% CO (CO25, CO50 and CO100 groups respectively). Body weight, specific growth rate and feed efficiency were significantly affected by the replacement of FO with CO. The final body weight of the CO100 group was significantly lower than that of the FO (control) and CO50 groups. Furthermore, the CO50 group showed significantly higher feed efficiency than the FO group during the first 4 weeks (long photoperiod and high water temperature conditions). Increasing dietary CO increased serum glucose and triglyceride concentrations in a dose‐dependent manner. Regarding the proximate composition of fillet and liver, only crude protein in fillet was affected by the replacement of FO with CO. Fatty acid compositions of fillet and liver were correlated to the amount of CO in each diet. The activities of the metabolic enzymes phosphofructokinase and alanine aminotransferase were lowest in the CO25 and CO50 groups respectively. Thus, FO replacement with CO affected growth performance, serum components, fatty acid composition and the activity of metabolic enzymes. Overall, results obtained in the present study suggest that partial replacement (50%) of FO with CO is effective and might increase juvenile yellowtail growth under long photoperiod and/or high water temperature conditions.