Three polycentric rumen fungi, LL, LC2 and Ruminomyces elegans (C2), isolated from the rumen of cattle were grown in six culture media. LL and LC2 were morphologically similar. Their characteristics resembled those of Orpinomyces and Neocallimastix joyonii, and they grew well and produced sporangia after 3-4 d growth in all the media. R. elegans differed morphologically from LL and LC2, but although it also grew well in all media, abundant sporangia occurred only after 2-3 d growth in media containing cellulose. Undifferentiated sporangia were produced by all three isolates; differentiation of the sporangia did not occur in the spent growth media. However, if thalli possessing recently-formed sporangia were transferred to, or flooded with, fresh liquid medium or rumen fluid, zoosporogenesis and liberation of zoospores occurred within 17-20 min for isolates LL and LC2 and 30 min for R. elegans. Procedures for inducing zoosporogenesis by polycentric anaerobic fungi are described.
A 45-year-old Malay lady who presented with intermittent abdominal pain and a left hypochondrial mass was found to have a 10 x 8 x 5 cm duodenal tumour without local invasion at laparotomy. En bloc resection of the tumour with adequate margin of clearance was done and histopathological diagnosis of low grade leiomyosarcoma was made. Fourteen months later, she returned with multiple metastases in the liver and needed palliative chemotherapy for pain relief. Duodenal leiomyosarcomata are very rare tumour. Their prognostic indicators include biological grading, tumour size and presence of metastases. Recognition of its high malignant potential calls for close surveillance calls even after apparent curative surgery.
Isozymes of 23 cultures of the anaerobic rumen fungi and seven cultures of aerobic chytridiomycete fungi were analysed by PAGE. A total of 14 isozyme loci were successfully typed by PAGE. They were peptidase A & C-1, peptidase A & C-2, peptidase D-1, peptidase D-2, malate dehydrogenase-1, malate dehydrogenase-2, esterase-1, esterase-2, malic enzyme-1, malic enzyme-2, isocitrate dehydrogenase, shikimate dehydrogenase, phosphoglucomutase and 6-phosphogluconate dehydrogenase. Isozyme analysis can be used for studying the genetic relationships among the different anaerobic rumen fungi and the aerobic chytridiomycete fungi and the isozyme characteristics can serve as additional taxonomic criteria in the classification of the anaerobic rumen fungi. A dendrogram based on the isozyme data demonstrated that the anaerobic rumen fungi formed a cluster, indicating a monophyletic group, distinctly separated from the aerobic chytridiomycete fungi. Piromyces communis and P. minutus showed a close relationship but P. spiralis showed a more distant relationship to both P. communis and P. minutus. Piromyces as a whole was more related to Caecomyces than to Neocallimastix. Orpinomyces was also found to be more related to Piromyces and Caecomyces than to Neocallimastix. Orpinomyces intercalaris C 70 from cattle showed large genetic variation from O. joyonii, indicating that it is a different species.
Voluntary food intake, digestibility and water turnover were determined in adult Malaysian lesser mouse-deer (Tragulus javanicus) given unlimited access to lundai foliage (Sapium baccatum). Daily dry matter (DM) intake was 42.4 g/kg metabolic live mass (M0.73) or 3.7% M. Digestible energy intake was 853 kJ/day (571 kJ metabolisable energy per M0.73), calculated to be used with 79% efficiency. Apparent digestibility (%) of organic matter was 83.8, crude fibre 63.7, acid detergent fibre 60.5, neutral detergent fibre 72.1 and crude protein 65.0. Urinary excretion of the purine derivative, allantoin, was 0.05 mg/g digestible DM intake suggesting rumen microbial yield efficiency may be lower than in other ruminant species. Total water intake was 182 ml/M0.82. The body-water content of the fed mouse-deer, from tritiated water dilution, was 77% M, consistent with a very lean carcass. Turnover of body water was 17% per day. The mouse-deer produced relatively dry, well-defined faecal pellets.
Isolates of anaerobic fungi obtained from the rumen, duodenum and faeces of sheep were identified as Piromyces mae based on their morphological characteristics observed using light microscopy. There was no significant morphological variation among the isolates of P. mae from the rumen, duodenum and faeces. Isozymes of 12 isolates of P. mae (one each from the rumen, duodenum and faeces from 4 different sheep) were analysed by PAGE. A total of 12 isozymes were studied and 5 isozyme loci were successfully typed. They were malic enzyme, malate dehydrogenase, shikimate dehydrogenase, alpha-esterase and beta-esterase. All the isolates of P. mae regardless of whether they were from the rumen, duodenum or faeces or from different animals produced very similar isozyme banding patterns for each of the enzyme systems. The similar isozyme profiles of the isolates indicate that they are of the same species although they exist in different regions of the alimentary tract.
Isotricha jalaludinii n. sp. found in the rumen of lesser mouse deer, Tragulus javanicus, in Malaysia was described and illustrated. This new species is characterized by the location and direction of the vestibulum, shape of the macronucleus, and absence of a dent at the vestibular opening. The presence of single peculiar isotrichid species in the rumen of mouse deer, which is recognized as one of the most primitive ruminants, suggests that the isotrichid ciliates similar to I. jalaludinii and Isotricha intestinalis were established at a fairly early period during the evolution of ruminants.
A total of 46 Lactobacillus isolates obtained from chicken intestine were assessed on their ability to adhere to the chicken ileal epithelial cell (IEC) in vitro. Twelve out of the 46 isolates showed moderate to good ability to adhere to the IEC. Temperature (between 4 degrees C and 42 degrees C) did not affect attachment. Incubation (contact) time of 30 min was found to be insufficient for the attachment of bacteria to the IEC, but contact time beyond 1 h did not increase this ability. The pH values (4-7) of the suspending buffer did not have any significant effect on the attachment of bacteria to the IEC, but at pH 8 it was reduced significantly (P < 0.05).
Single strains of Lactobacillus acidophilus and Lact. fermentum, isolated from chicken intestine, were used to study in vitro interactions with Salmonella enteritidis, Salm. pullorum or Salm. typhimurium in an ileal epithelial cell (IEC) radioactive assay. Exclusion, competition and displacement phenomena were investigated by respectively incubating (a) lactobacilli and IEC together, prior to addition of salmonellae, (b) lactobacilli, IEC and salmonellae together, and (c) salmonellae and IEC, followed by the lactobacilli. Lactobacilli were selected for study because of their strong ability to adhere to IEC and poor aggregation with salmonellae. The results demonstrated that Lact. acidophilus significantly reduced (P < 0.05) the attachment of Salm. pullorum to IEC in the tests for exclusion and competition, but not in the displacement tests. Lactobacillus fermentum was found to have some ability to reduce the attachment of Salm. typhimurium to IEC under the conditions of exclusion (P < 0.08), competition (P < 0.09), but not displacement. However, both Lact. acidophilus and Lact. fermentum were unable to reduce the adherence of Salm. enteritidis to IEC under any of the conditions.
Twelve Lactobacillus strains isolated from chicken intestine, which demonstrated a strong and moderate capacity to adhere to the ileal epithelial cells in vitro, were used to investigate their inhibitory ability against five strains of salmonella, i.e. Salmonella enteritidis 935/79, Salm. pullorum, Salm. typhimurium, Salm. blockley and Salm. enteritidis 94/448, and three serotypes of Escherichia coli, viz. E. coli O1:K1, O2:K1 and O78:K80. The results showed that all the 12 Lactobacillus isolates were able to inhibit the growth of the five strains of salmonella, and the three strains of E. coli in varying degrees. Generally, they were more effective in inhibiting the growth of salmonella than E. coli. Inhibition of the pathogenic bacteria was probably due to the production of organic acids by the Lactobacillus isolates.
733 senior civil servants comprising 520 males and 213 females with a mean age of 44 years (range 25-56 years) were screened for their health status. The sample population comprised of 67.9% Malays, 22.5% Chinese, 9.1% Indians and 0.4% other ethnic origins. The subjects' medical histories were recorded and a full medical examination including anthropometric measurements (weight, height, waist and hip circumference), blood biochemistry and urine analysis, chest X-ray and electrocardiograms were done. The results obtained showed that 36.0% of the study population were overweight with 6.5% being obese. Of this 32.0% had central obesity. 15.2% of the subjects had systolic hypertension (systolic BP > or = 140 mmHg) whilst 27.6% had diastolic hypertension (diastolic BP > or = 90 mmHg). Hyperlipidaemia was common, with 75.2% subjects having raised cholesterol, 19.9% raised triglycerides, 50.2% raised LDL-C, 74.6% raised TC:HDL-C and 26.6% raised LDL:HDL-C. An elevated blood glucose was found in 8.4% subjects, whilst urine sugar was detected in only 2.6%, and a raised uric acid was found in 2.8% subjects. The prevalence of hypertension, raised blood glucose and hyperlipidaemia increased with age with more males affected than females. Although hypercholesterolaemia appeared more frequently amongst the Malays, the Indians, by comparison had the highest prevalence for a raised LDL:HDL-C ratio, a reflection of the increase in LDL-C and a concomitant decrease in HDL-C. The latter findings indicate that the Indians are at greater risk for the development of coronary heart disease than the Chinese and Malays. In addition, the mean levels of serum cholesterol found in this study seemed to have exceeded the levels found in populations in the industrialised countries such as the USA. There is thus an urgent need for more public health campaigns aimed at the reduction and control of such coronary risk factors.
Two Lactobacillus isolates, Lact. acidophilus I 26 and Lact. fermentum I 25, were selected, based on their poor aggregation with Escherichia coli and strong ability to adhere to ileal epithelial cells (IEC), to study in vitro interactions with E. coli O1:K1, O2:K1 and O78:K80 in an IEC radioactive-assay under the conditions of exclusion (lactobacilli and IEC, followed by the addition of E. coli), competition (lactobacilli, IEC and E. coli together) and displacement (E. coli and IEC, followed by the addition of lactobacilli). The results indicated that Lact. acidophilus I 26 and Lact. fermentum I 25 could not significantly reduce the attachment of E. coli O1:K1, O2:K1 and O78:K80 to IEC under the three conditions tested in vitro, except that the attachment of E. coli O1:K1 was slightly reduced by Lact. fermentum I 25 in the test for competition.
A study was conducted to determine the effects of adherent Lactobacillus culture on growth performance, intestinal microbial population, and serum cholesterol level of broilers. Four dietary treatments, consisting of the basal diet (control), basal diet + 0.05, 0.10, or 0.15% Lactobacillus culture (LC), were fed to 2,000 Arbor Acres broiler chicks from 1 to 42 d of age (DOA). The chicks were randomly assigned to 40 cages (50 chicks per cage, 10 cages per diet). The experimental period was 42 d. Body weights and feed to gain ratio were measured at 21 and 42 DOA. The intestinal microbial populations and serum cholesterol levels were determined at 10, 20, 30, and 40 DOA. The results showed that body weights and feed to gain ratios were improved significantly (P < 0.05) when compared to control broilers for broilers fed diets containing 0.05 or 0.10% LC, but not 0.15% LC, at 21 and 42 DOA. Coliform counts in the cecum of birds receiving 0.05% LC at 10, 20, and 30 DOA, and 0.10% at 10 and 20 DOA were significantly lower (P < 0.05) than those of the control birds. The total aerobes, total anaerobes, lactobacilli, and streptococci in the small intestines and ceca of the control birds were not significantly different from those of the treated groups. Serum cholesterol levels were significantly lower (P < 0.05) in broilers fed the three diets containing LC at 30 DOA, and in the birds fed 0.05 or 0.10% LC at 20 DOA.
Twelve Lactobacillus strains isolated from chicken intestine were used to investigate acid and bile tolerance in vitro. Ten out of the 12 strains were slightly affected by 0.3% bile salts, showing a delay of growth (d) of 0.6-37.2 min compared with growth in control cultures. Two strains were not affected by the bile salts. Of the 12 strains, seven could be arbitrarily classified as resistant (d < 15 min) and five as tolerant (15 min < d < or = 40 min). Lactobacillus strains from the caecum showed better tolerance to acid than those from the ileum. Generally, the survival of the ileal strains was very low at pH 1.0 and 2.0, and moderate at pH 3.0. In contrast, caecal Lactobacillus strains could survive at pH 1.0 for up to 2 h of incubation; growth was moderate at pH 2.0 and good at pH 3.0 and 4.0.
A total of 31 strains of Vibrio cholerae O1 (10 from outbreak cases and 7 from surface water) and non-O1 (4 from clinical and 10 from surface water sources) isolated between 1993 and 1997 were examined with respect to presence of cholera enterotoxin (CT) gene by PCR-based assays, resistance to antibiotics, plasmid profiles and random amplified polymorphic DNA (RAPD) analysis. All were resistant to 9 or more of the 17 antibiotics tested. Identical antibiotic resistance patterns of the isolates may indicate that they share a common mode of developing antibiotic resistance. Furthermore, the multiple antibiotic resistance indexing showed that all strains tested originated from high risk contamination. Plasmid profile analysis by agarose gel electrophoresis showed the presence of small plasmids in 12 (7 non-O1 and 5 O1 serotypes) with sizes ranging 1.3-4.6 MDa. The CT gene was detected in all clinical isolates but was present in only 14 (6 O1 serotype and 8 non-O1 serotype) isolates from environmental waters. The genetic relatedness of the clinical and environmental Vibrio cholerae O1 and non-O1 strains was investigated by RAPD fingerprinting with four primers. The four primers generated polymorphisms in all 31 strains of Vibrio cholerae tested, producing bands ranging from < 250 to 4500 bp. The RAPD profiles revealed a wide variability and no correlation with the source of isolation. This study provides evidence that Vibrio cholerae O1 and non-O1 have significant public health implications.
1. Hubbard x Hubbard (HH) and Shaver x Shaver (SS) chicks given a dietary supplement of either 50 mg/kg oxytetracycline (OTC) or 1 g/kg Lactobacillus culture (LC) were exposed to 36 +/- 1 degrees C for 3 h daily from day (d) 21 to 42. 2. Prior to heat treatment, body weight (d 21) and weight gain (d 1 to d 21) of OTC and LC birds were greater than those fed the control diet. Chicks given LC had the best food efficiency followed by OTC and control birds during d 1 to d 21. Body weight (d 1 and d 21) and weight gain (d 1 to d 21) were greater for HH tlhan SS chicks. 3. After 3 weeks of heat exposure, birds receiving the LC diet had greater body weight and weight gain, higher food intake and lower food efficiency than OTC and control chicks. 4. Antibody production against Newcastle discase vaccine on d 21 was not affected by strain or diet. On d 42, while diet had negligible effect on this variable among the SS broilers, HH birds fed LC had higher antibody production than those on the control diet. 5. Neither strain nor diet had a significant effect on mortality.
Heat production (HP) of male and female mouse deer during eating, standing and sitting was determined using the open circuit respiration chamber (RC). The time taken for similar activities was also determined in an outdoor enclosure (OD). The animals were fed kangkong (Ipomoea aquatica), sweet potato (Ipomoea batatas) and rabbit pellet ad libitum. Male mouse deer consumed more dry matter (DM), organic matter (OM) and gross energy (GE) than female. The time for each activity of male and female mouse deer kept in RC and OD was similar. The average time spent in RC and OD for both male and female, respectively, for sitting (956 and 896 min/day) was significantly (P<0.01) longer than standing (463 and 520 min/day) and eating (21 and 24 min/day). Heat production for male and female mouse deer, respectively, during eating was the highest (0.44 and 0.43 kJ/kg W(0.75)/min) followed by standing (0.37 and 0.33 kJ/kgW(0.75)/min) and sitting (0.26 and 0.26 kJ/kg W(0.75)/min). The difference in HP per min during standing between male and female was significant (P<0.05). The HP for 08.00-14.00 h and 14.00-20.00 h periods were higher than 20.00-02.00 h and 02.00-08.00 h periods. The overall HP for males during 08.00-14.00 h and 14.00-20.00 h periods were significantly (P<0.05) higher (114.8 and 119.2 kJ/kg W(0.75)) than female (107.5 and 110.4 kJ/kg W(0.75)), respectively.
Five strains of phytase-producing, gram-negative, non-spore-forming, non-motile, small, stout, rod-shaped, strictly anaerobic, fermentative bacteria were isolated from the rumens of cattle in Malaysia. All five strains had morphological, physiological and biochemical features in common. Although these strains had many physiological and biochemical characteristics that were identical to those of the Mitsuokella multacida type strain (ATCC 27723T), they could be distinguished from this species by means of the following characteristics: a smaller cell size (1.2-2.4 microm long and 0.6-0.8 microm wide); a lower final pH value (3.8-4.0) in peptone/yeast extract/glucose broth; inhibition by 0.001% brilliant green; insensitivity to kanamycin (100 microg ml(-1)) and penicillin (10 microg ml(-1)); a higher optimum growth temperature (approx. 42 degrees C); the ability to grow at 45 and 47 degrees C; the ability to ferment glycerol, sorbitol and amidon; and the inability to ferment mannitol, rhamnose, D-tagatose and melezitose. The G+C content of the type strain (M 9T) of these five strains was 56.9 mol%. Analysis of the 16S rRNA gene sequence of type strain M 9T indicated that the strain falls within the genus Mitsuokella. The sequence similarity between type strain M 9T and Mitsuokella multacida was 98.7%. The DNA-DNA relatedness between type strain M 9T and Mitsuokella multacida type strain DSM 20544T (= ATCC 27723T) was 63.8%, indicating that, in spite of a high level of similarity for the 16S rRNA gene sequence, type strain M 9T is independent of Mitsuokella multacida at the species level. On the basis of these results, a new species, Mitsuokella jalaludinii sp. nov., is proposed for these strains. The type strain is M 9T (= DSM 13811T = ATCC BAA-307T).
We evaluated the efficacy of supplementation of active Mitsuokella jalaludinii culture (AMJC) on the growth performance, nutrient use, and mineral concentrations in tibia bone and plasma of broiler chickens fed corn-soybean meal diets. Dietary treatments included low-nonphytate P (NPP) feed (containing 0.24% and 0.232% NPP for chicks from 1 to 21 and 22 to 42 d of age, respectively), low-NPP feed added with different levels of AMJC (equivalent to 250, 500, 750, and 1,000 U phytase/kg of feed), and normal-NPP feed (containing 0.46 and 0.354% NPP for chicks from 1 to 21 and 22 to 42 d of age, respectively). Supplementation of AMJC to low-NPP feed increased (P < 0.05) weight gain and feed intake and decreased (P < 0.05) feed:gain ratio of chickens during the whole experiment (Days 1 to 42). Supplementation of AMJC increased (P < 0.05) the AME value, digestibility of DM and CP, and retention of P, Ca, and Cu. Mn retention in broilers was only increased (P < 0.05) by AMJC supplementation from 18 to 20 d of age, and Zn retention was improved (P < 0.05) only at a high level of AMJC (equivalent to 1,000 U phytase/kg of feed) supplementation. Chicks fed low-NPP feed added with AMJC had similar tibia ash percentages as those fed the normal-NPP diet. Generally, supplementing AMJC to low-NPP feed increased (P < 0.05) Ca, decreased significantly (P < 0.05) Mn and Cu, but did not affect Zn and P concentrations in tibia ash. Supplementing AMJC also increased (P < 0.05) plasma P but had no effect on plasma Ca or Mn. Plasma Zn concentration was increased only when a high level of AMJC (equivalent to 1,000 U phytase/kg of feed) was used. In conclusion, AMJC supplementation to low-NPP feed improved growth performance; AME value; digestibility of CP and DM; use of Ca, P, and Cu; and bone mineralization.
The effects of pH, temperature, phytate, glucose, phosphate and surfactants on the phytase production of Mitsuokella jalaludinii, a new bacterial species from the rumen of cattle, were evaluated.
The effects of different carbon and nitrogen sources on phytase production by Mitsuokella jalaludinii were evaluated and the optimization of rice bran (RB) and soybean milk (SM) concentrations in the medium for phytase production was also determined.