Influences of short-term pre-slaughter dietary manipulation in sheep and goats on pH and microbial loads of gastrointestinal tract

Sheep (BW = 39.9 kg, n = 16) and goats (BW = 32.8 kg, n = 16) were used in a completely randomized design to determine the effect of short-term pre-slaughter diet and feed deprivation (FD) time on pH and microbial loads in the gastrointestinal tract (GIT) contents. In a 2 × 2 × 2 factorial treatment arrangement, the main effects of species, diet, and FD time prior to slaughter and their interactions were studied. Animals were fed either a hay or concentrate diet for 4 d and then feed deprived for either 12 or 24-h prior to slaughter. The pH of rumen and colon contents as well as weight of GIT was measured. The contents of rumen and rectum were also sampled for microbial analysis. The GIT of sheep (1.82 kg) was heavier (P < 0.05) than that of goats (1.46 kg). The 12-h FD group (1.74 kg) had a higher (P < 0.05) GIT weight than the 24-h FD group (1.53 kg). Hay-fed animals had higher (P < 0.05) rumen (7.08 vs. 6.43) and colon pH values (7.02 vs. 6.56) than those of the concentrate-fed animals. The 24-h FD group (3.39 ± 0.272 log₁₀CFU/g) contained more (P < 0.05) Escherichia coli in the rumen than did the 12-h FD (2.17 ± 0.272 log₁₀CFU/g) group. The concentrate-fed animals (3.49 ± 0.289 log₁₀CFU/g) had higher (P < 0.05) coliform counts in the rumen than the hay-fed animals (2.43 ± 0.289 log₁₀CFU/g). The 24-h FD group (3.42 ± 0.289 log₁₀CFU/g) had a higher (P < 0.05) concentration of coliform than did the 12-h FD group (2.50 ± 0.289 log₁₀CFU/g). The 24-h FD group (3.31 ± 0.259 log₁₀CFU/g) also had higher (P < 0.05) Enterobacteriaceae counts in the rumen than did in the 12-h FD group (2.47 ± 0.259 log₁₀CFU/g). Goats (5.71 ± 0.158 log₁₀CFU/g) had lower (P < 0.05) total plate counts in the rumen compared to sheep (6.27 ± 0.158 log₁₀CFU/g). The concentrate-fed animals had higher (P < 0.05) E. coli (6.44 vs. 4.01 ± 0.468 log₁₀CFU/g), total coliform (6.74 vs. 4.16 ± 0.469 log₁₀CFU/g), Enterobacteriaceae (6.93 vs. 3.83 ± 0.651 log₁₀CFU/g), and total plate counts (7.79 vs. 7.28 ± 0.170 log₁₀CFU/g) in the rectum than the hay-fed animals. The results indicate that microbial loads in the GIT of small ruminants may be reduced by either feeding hay for 4 d or depriving feed for 12-h prior to slaughter.     

Comparison of the archaeal community in the fermentative compartment and faeces of the cow and the rabbit

The archaeal community in the fermentative compartment and faeces of the cow and the rabbit were compared by analysis capillary electrophoresis single-stranded conformation polymorphism (CE-SSCP) profiles of 16S rRNA genes. Ruminal and faecal contents were sampled in five cows for three weeks. Hard and soft faeces were collected in 14 rabbits for three consecutive weeks and cæcal contents were sampled in the third week. The archaeal community differed according to the host species (ANOSIM-R = 0.53 and 0.72 respectively for the comparison of the fermentative compartments and faeces; P < 0.001) and to the location within the digestive tract of both species (ANOSIM-R = 0.37, 0.52 respectively for the cow and the rabbit; P < 0.001). In both species, the archaeal community of the digestive tract was stable over weeks and varied very little between individual animals. The structure (NS) and the richness index (9.9 ± 2.7, 10.1 ± 3.1 respectively, NS) of the archaeal community were similar for the cæcal content and the soft faeces which permitted to use the latter as a representative indicator.     

Copulation duration of bumblebee Bombus terrestris (Hymenoptera: Apidae): Impacts on polyandry and colony parameters

The influence of copulation duration on mating frequency and colony development were studied in Bombus terrestris (Hymenoptera: Apidae). Copulation time was recorded in transparent plastic boxes and was manipulated by separating mating pairs. Mean copulation duration was found to be 30.0 ± 8.0 (mean ± se) minutes and most matings lasted 20 to 40 min. When queens were only allowed to mate for 2 to 5 min, the chance that they would accept a second mating was 7.2 ± 5.0 % (mean ± se). Incompletely mated queens delayed to initiate colonies but they did not show significant difference from fully mated queens in production of new queens and males. This study shows that colony development was not affected by short copulation duration.     

A novel medium for the production of cephamycin C by Nocardia lactamdurans using solid-state fermentation

In this study, Nocardia lactamdurans NRRL 3802 was explored for the first time for production of cephamycin C by using solid-state fermentation. The effects of various substrates, moisture content, inoculum size, initial pH of culture medium, additional nitrogen source and amino acids were investigated for the maximum production of cephamycin C by N. lactamdurans NRRL 3802 in solid-state fermentation. Subsequently, selected fermentation parameters were further optimized by response surface methodology (RSM). The soybean flour as a substrate with moisture content of 65%, initial pH of culture medium of 6.5 and inoculum size of 10⁹ CFU/ml (2 × 10⁸ CFU/gds) at 28 ± 2 °C after 4 days gave maximum production of 15.75 ± 0.27 mg/gds of cephamycin C as compared to 8.37 ± 0.23 mg/gds before optimization. Effect of 1,3-diaminopropane on cephamycin C production was further studied, which further increased the yield to 27.64 ± 0.33 mg/gds.     

Enzymatic properties of a bee venom serine protease from the bumblebee Bombus ignitus

We previously reported that bumblebee (Bombus ignitus) venom serine protease (Bi-VSP) acts as a prophenoloxidase-activating factor in arthropods and a fibrin(ogen)olytic enzyme in mammals. In the present study, we characterized the enzymatic properties of Bi-VSP purified from B. ignitus venom. The 34-kDa active form of Bi-VSP was purified from the venom of B. ignitus worker bees. Glycoprotein staining showed that approximately 20% of the total molecular mass of Bi-VSP is due to carbohydrate moieties. Bi-VSP had an optimal pH and temperature of pH 9.0 and 40 °C, respectively, and was stable at 50 °C for at least 10 min. Bi-VSP activity decreased abruptly below pH 6.0, indicating that Bi-VSP activity is almost completely inhibited at pH 5.4 of B. ignitus venom. The protease activity of Bi-VSP was strongly inhibited by typical serine protease inhibitors such as phenylmethanesulfonyl fluoride, leupeptin, and soybean trypsin inhibitor.     

Intestinal and immunological effects of daily oral administration of Lactobacillus salivarius CECT5713 to healthy adults

There is an increasing interest in the intestinal and immunological effects of probiotics. The aim of the present study is to evaluate the tolerance and beneficial effects in healthy adults of the strain, Lactobacillus salivarius CECT5713 isolated from breast milk. A phase II, randomized, double-blinded, placebo-controlled human clinical trial was carried out in 40 healthy adults. The Probiotic group received a daily dose of 2 × 10⁸ CFU of L. salivarius CECT5713 in capsules during 4 weeks while volunteers of the control received only a placebo. Gastrointestinal and immunological parameters were analyzed. Results showed that L. salivarius CECT5713 was well tolerated and no adverse effects were detected. Consumption of the probiotic strain increased fecal lactobacilli counts (7.9 ± 0.1 vs. 7.05 ± 0.2 CFU/g feces, P = 0.001). Also, an improvement in the frequency of defecation (P = 0.04) was observed. Probiotic treatment induced significantly the percentage of NK cells and monocytes, as well as the plasmatic levels of immunoglobulins M, A and G, and the regulatory cytokine IL-10 (72.3 ± 11.7 in probiotic group vs. 27.3 ± 6.4 pg/mL in control group, P < 0.01). Thus, it can be concluded that daily administration of L. salivarius CECT5713 to healthy adults is safe and improve gut microbiota and different parameters related to immune response.     

Nutritional value of Chlorella vulgaris: Effects of ultrasonication and electroporation on digestibility in rats

Three processed products derived from the green algae C. vulgaris were investigated: (1) spray-dried only (S-DA); (2) spray-dried and electroporated (ES-DA); (3) spray-dried; ultrasonicated treated (US-DA). A nitrogen-balance study was performed. Male growing Wistar rats, housed separately in metabolism cages, were fed the three algal products as the sole protein source at 150 mg N per 100 g of body weight. A control group of rats was fed with casein at a level to give the same protein nitrogen intake. The coefficients of total intestinal tract apparent crude protein digestibility for the different C. vulgaris products were: S-DA = 0.47 ± 0.127% (mean ± S.D.), ES-DA = 0.44 ± 0.075%, US-DA = 0.57 ± 0.137%. Protein efficiency ratio was 1.4 ± 0.3, 1.0 ± 0.5 and 2.1 ± 0.3, respectively. N-balance was 41.86 ± 32.8 mg, 31.3 ± 17.3 mg and 66.7 ± 30.1 mg, respectively. The biological value was 93 ± 9.5%, 93.6 ± 10%, and 101 ± 5%, respectively. The coefficient of total intestinal tract apparent crude protein digestibility and biological value of C. vulgaris was enhanced by ultrasonic treatment and reduced by electroporating, thus ultrasonication may be a helpful technological process in practical processing of green algae in food industry.     

Stability constants and multinuclear NMR measurements of phosphonic acid derivatives with aluminum in aqueous solutions

We are reporting the stability constants of the different complexes between phosphonoacetic acid (PAA), phosphonoformic acid (PFA), aminomethylphosphonic acid (AMPA), aminoethylphosphonic acid (AEPA) and methylenediphosphonic acid (MDP) with the aluminum metal ion in aqueous solutions. (In this study the term aluminum is reserved for the 3+ ion.) The affinity of the phosphonic acid derivatives to chelate aluminum has been tested by potentiometric titrations with I = 0.10 M KNO₃ at 25 ± 0.1 °C. The proposed aluminum–ligand complex structures have been confirmed by ³¹P NMR, ¹³C NMR, and ²⁷Al NMR experiments. Both PAA and PFA formed simple one to one complexes. The respective values for PAA are log β₁₁₁ = 13.57, log β₁₁₀ = 10.58, and log β₁₁₋₁ = 5.84. The respective values for PFA are log β₁₁₂ = 15.24, log β₁₁₁ = 13.11, and log β₁₁₀ = 6.88. In contrast to PAA and PFA, the major species formed with AMPA and AEPA consist of a series of dimeric complexes. The ³¹P NMR spectra of these complexes indicate that the amine groups do not co-ordinate to aluminum and remain protonated. In addition to these dimeric complexes, a monoprotonated monomer of Al–AMPA also has been detected. The ²⁷Al NMR experiments indicated that the aluminum is hexacoordinated in all the complexes in this study and the hydroxide ion did not remove aluminium from its complexes. Among the ligands studied, PAA and PFA were able to solubilize aluminum at physiological pH. A comparison between the acidities and the chelating properties of the ligands used is presented.     

Effect of probiotic L. plantarum IS-10506 and zinc supplementation on humoral immune response and zinc status of Indonesian pre-school children

A 90-day randomized, double-blind, placebo-controlled, pre-post trial was conducted in four groups of Indonesian children aged 12–24 months: placebo, probiotic, zinc, and a combination of probiotic and zinc (n = 12 per group). Microencapsulated Lactobacillus plantarum IS-10506 of dadih origin was supplemented at a dose of 10¹⁰ CFU/day as a probiotic. Zinc was supplemented as 20 mg zinc sulfate monohydrate (8 mg zinc elemental). Blood and stool samples were collected at baseline and at the end of the study period. Fecal sIgA was assessed by ELISA and serum zinc concentrations by ICP-MS. Fecal sIgA increased significantly in the probiotic group (30.33 ± 3.32 μg/g; p < 0.01) and in the combination probiotic and zinc group (27.55 ± 2.28 μg/g; p < 0.027), as compared with the placebo group (13.58 ± 2.26 μg/g). Changes in serum zinc concentrations in the combination probiotic and zinc group showed the highest elevation at the end of the study period. A combination of probiotic L. plantarum IS-10506 at a dose of 10¹⁰ CFU/day and 8 mg of elemental zinc supplementation showed a potential ability to improve the zinc status of pre-school children. Taken together, supplementation with the probiotic L. plantarum IS-10506 and zinc for 90 days resulted in a significantly increased humoral immune response, as well as improved zinc status, in young children.     

Gastro-intestinal handling of water and solutes in three species of elasmobranch fish,the white-spotted bamboo shark,Chiloscyllium plagiosum,little skate,Leucoraja erinacea and the clear nose skate Raja eglanteria

The present study reports aspects of GI tract physiology in the white-spotted bamboo shark, Chiloscyllium plagiosum, little skate, Leucoraja erinacea and the clear nose skate, Raja eglanteria. Plasma and stomach fluid osmolality and solute values were comparable between species, and stomach pH was low in all species (2.2 to 3.4) suggesting these elasmobranchs may maintain a consistently low stomach pH. Intestinal osmolality, pH and ion values were comparable between species, however, some differences in ion values were observed. In particular Ca²⁺ (19.67 ± 3.65 mM) and Mg²⁺ (43.99 ± 5.11 mM) were high in L. erinacea and Mg²⁺ was high (130.0 ± 39.8 mM) in C. palgiosum which may be an indication of drinking. Furthermore, intestinal fluid HCO₃^? values were low (8.19 ± 2.42 and 8.63 ± 1.48 mM) in both skates but very high in C. plagiosum (73.3 ± 16.3 mM) suggesting ingested seawater may be processed by species-specific mechanisms. Urea values from the intestine to the colon dropped precipitously in all species, with the greatest decrease seen in C. plagiosum (426.0 ± 8.1 to 0 mM). This led to the examination of the molecular expression of both a urea transporter and a Rhesus like ammonia transporter in the intestine, rectal gland and kidney in L. erinacea. Both these transporters were expressed in all tissues; however, expression levels of the Rhesus like ammonia transporter were orders of magnitude higher than the urea transporter in the same tissue. Intestinal flux rates of solutes in L. erinacea were, for the most part, in an inward direction with the notable exception of urea. Colon flux rates of solutes in L. erinacea were all in an outward direction, although absolute rates were considerably lower than the intestine, suggestive of a much tighter epithelia. Results are discussed in the context of the potential role of the GI tract in salt and water, and nitrogen, homeostasis in elasmobranchs.     

Angiotensin II-induced activation of c-Ret signaling is critical in ureteric bud branching morphogenesis

The renin-angiotensin system (RAS) plays a critical role in ureteric bud (UB) and kidney morphogenesis. Mutations in the genes encoding components of the RAS cause a spectrum of congenital abnormalities of the kidney and urinary tract (CAKUT). However, the mechanisms by which aberrations in the RAS result in CAKUT are poorly understood. Given that c-Ret receptor tyrosine kinase (RTK) is a major inducer of UB branching, the present study tested the hypothesis that angiotensin (Ang) II-induced activation of c-Ret plays a critical role in UB branching morphogenesis. E12.5 mice metanephroi were grown for 24 h in the presence or absence of Ang II, Ang II AT₁ receptor (AT₁R) antagonist candesartan, phosphatidylinositol 3-kinase (PI3 K) inhibitor LY294002 or ERK1/2 inhibitor PD98059. Ang II increased the number of UB tips (61 ± 2.4 vs. 45 ± 4.3, p < 0.05) compared with control. Quantitative RT-PCR analysis demonstrated that Ang II increased c-Ret mRNA levels in the kidney (1.35 ± 0.05 vs. 1.0 ± 0, p < 0.01) and in the UB cells (1.28 ± 0.04 vs. 1.0 ± 0, p < 0.01) compared to control. This was accompanied by increased Tyr¹⁰⁶²Ret phosphorylation by Ang II (5.5 ± 0.9 vs. 1.8 ± 0.4 relative units, p < 0.05). In addition, treatment of UB cells with Ang II (10^?5 M) increased phosphorylation of Akt compared to control (213 ± 16 vs. 100 ± 20%, p < 0.05). In contrast, treatment of metanephroi or UB cells with candesartan decreased c-Ret mRNA levels (0.72 ± 0.06 vs. 1.0 ± 0, p < 0.01; 0.68 ± 0.07 vs. 1.0 ± 0, p < 0.05, respectively) compared with control. Ang II-induced UB branching was abrogated by LY294002 (24 ± 2.6 vs. 37 ± 3.0, p < 0.05) or PD98059 (33 ± 2.0 vs. 48 ± 2.2, p < 0.01). These data demonstrate that Ang II-induced UB branching depends on activation of Akt and ERK1/2. We conclude that cross-talk between the RAS and c-Ret signaling plays an important role in the development of the renal collecting system.     

Peel of araticum fruit (Annona crassiflora Mart.) as a source of antioxidant compounds with α-amylase, α-glucosidase and glycation inhibitory activities

Annona crassiflora Mart., whose fruit is popularly known as araticum, is a member of the Annonaceae family found in the Brazilian Cerrado. Although this plant has several medicinal uses, its bioactive molecules are not fully understood. A bioguided assay was performed to identify the main bioactive compounds of A. crassiflora fruit peel from the ethanol extract fractions with antioxidant capacity and α-amylase, α-glucosidase and glycation inhibitory activities. Ethyl acetate and n-butanol fractions showed, respectively, higher antioxidant capacity (DPPH IC₅₀ 1.5 ± 0.1 and 0.8 ± 0.1 μg mL⁻¹, ORAC 3355 ± 164 and 2714 ± 79 μmol trolox eq/g, and FRAP 888 ± 16 and 921 ± 9 μmol trolox eq/g) and inhibitory activities against α-amylase (IC₅₀ 4.5 ± 0.8 and 1.7 ± 0.3 μg mL⁻¹), α-glucosidase (IC₅₀ 554.5 ± 158.6 and 787.8 ± 140.6 μg mL⁻¹) and glycation (IC₅₀ 14.3 ± 3.3 and 16.0 ± 4.2 μg mL⁻¹), and lower cytotoxicity, compared to the other fractions and crude ethanol extract. The HPLC-ESI-MS/MS analysis identified various biomolecules known as potent antioxidants, such as chlorogenic acid, (epi)catechin, procyanidins, caffeoyl-hexosides, quercetin-glucosides and kaempferol. The fruit peel of A. crassiflora, a specie from Cerrado, the Brazilian Savanna, provided a source of antioxidant compounds with properties to block carbohydrate digestive enzymes and formation of glycation products. Thus, there is potential to use the by-products of araticum in order to identify and isolate phytochemicals for application in nutraceutical supplements, food additives and pharmaceuticals products.     

Microbial quality of common carp and pikeperch fingerlings cultured in a pond fed with treated wastewater

A pond with a surface area of 0.94 ha and a maximum depth of 1.6 m, situated on land owned by a wastewater treatment plant in Olsztynek (Poland), was filled with water from underground springs in the pond bottom. Twice during the production season, the pond was fed with sewage which had been biologically treated in sequencing batch reactors (SBR). No commercial feed or medicinal products were applied to the pond and natural feed in the pond was the only source of nutrition for the fish. After the production season, pikeperch and carp fingerlings (aged 0+), weighed 8.4 ± 1.2 g fish^?1 and 62.1 ± 13.6 g fish^?1, respectively.The microbiological examinations included tests of the pond water and the contents of the digestive tract, skin mucus and the muscles of the fingerlings. In the water the total cell count, heterotrophic bacteria count at a temperature of 22 °C and 37 °C (HPC 22 °C and 37 °C), coliforms, fecal coliforms, Streptococcus faecalis, Salmonella sp. and Aeromonas hydrophila, were recorded. The same groups of bacteria were noted in the analyzed fish tissue without the total cell count. In addition, the antibiotic sensitivity of A. hydrophila, isolated from all the examined environments, was tested.The low count of all examined groups of bacteria in the pond water was noted. The common carp tissues were more contaminated than the pikeperch tissues, but the microbial quality of both species was satisfactory. A. hydrophila strains proved to be highly sensitive to the tested antibiotics. The results of this study indicate that a high microbiological quality in the fingerlings was achieved thanks to the seasonal feeding of pond water by treated sewage.     

Biochemical changes in the follicular fluid of the dominant follicle of high producing dairy cows exposed to heat stress early post-partum

High yielding dairy cows experience a negative energy balance (NEB) early post-partum and it was hypothesized that this may be aggravated under summer heat stress (HS) conditions. In this study, which was performed in Egypt, 20 Holstein cows were followed during summer (n = 10) and winter (n = 10) seasons. All cows were multiparous and kept at the same herd. Blood was sampled from each cow starting 1 week before the expected calving date and then at 1-week intervals until week 6 post-partum. From week 2 to 6 post-partum follicular fluid was collected through transvaginal follicular fluid aspiration at 6 days intervals. Ambient air temperature (AT) and relative humidity (RH) were recorded and temperature–humidity index (THI) was calculated as well. Respiration rate (RR), rectal temperature (RT), and body condition score (BCS) were recorded for each cow at the time of blood sampling. Concentrations of glucose, insulin like growth factor-1 (IGF-1), non-esterified fatty acids (NEFA), urea and total cholesterol (TC) were measured in each blood and follicular fluid sample. All the cows showed a significantly higher RR and RT in summer (95.5 ± 1.1 and 39.88 ± 0.06, respectively) than in winter (43.89 ± 0.61 and 38.94 ± 0.07, respectively) (P < 0.001). Body condition score loss during the early post-partum period was higher in summer than in winter (1.1 ± 0.07 vs. 0.85 ± 0.06 point, respectively) (P < 0.001). The average dominant follicle diameter was significantly lower in summer than in winter during the period of negative energy balance (11.6 ± 0.7 mm vs. 15.3 ± 1.2 mm, respectively) (P < 0.01). Under summer heat stress, the concentrations of glucose (2.98 ± 0.07 and 2.19 ± 0.04 mmol/L), IGF-1 (106.7 ± 2.9 and 99.0 ± 3.4 ng/ml) and TC (137.3 ± 5.3 and 62.2 ± 5.1 mg/dl) in blood and FF, respectively, were significantly lower than winter concentrations by (0.17 ± 0.03 mmol/L, P < 0.001 and 0.26 ± 0.06 mmol/L, P < 0.001), (12.3 ± 3.6 ng/ml, P < 0.001 and 9.0 ± 2.7 ng/ml, P < 0.001) and (20.7 ± 1.8 mg/dl, P < 0.001 and 7.3 ± 1.1 mg/dl, P < 0.01), respectively. However, the concentrations of NEFA (0.68 ± 0.14 and 0.22 ± 0.02 mmol/L) and urea (9.27 ± 0.34 and 9.96 ± 0.25 mmol/L) in blood and FF, respectively, were significantly higher in summer compared to winter (0.50 ± 0.08 mmol/L, P < 0.001 and 0.20 ± 0.02 mmol/L, P < 0.001) and (8.77 ± 0.23 mmol/L, P < 0.05 and 8.96 ± 0.29 mmol/L, P < 0.001), respectively, throughout the experimental period. The results of the present study indicate that heat stress early post-partum aggravates NEB in high yielding dairy cows, reduces BCS, dominant follicle diameter and alters the biochemical concentrations in the follicular fluid of the dominant follicle which may result in inferior oocyte and granulosa cell quality and hence poorer fertility.     

Use of Saccharum spontaneum (wild sugarcane) as biomaterial for cell immobilization and modulated ethanol production by thermotolerant Saccharomyces cerevisiae VS3

Saccharum spontaneum is a wasteland weed consists of 45.10 ± 0.35% cellulose and 22.75 ± 0.28% of hemicellulose on dry solid (DS) basis. Aqueous ammonia delignified S. spontaneum yielded total reducing sugars, 53.91 ± 0.44 g/L (539.10 ± 0.55 mg/g of substrate) with a hydrolytic efficiency of 77.85 ± 0.45%. The enzymes required for hydrolysis were prepared from culture supernatants of Aspergillus oryzae MTCC 1846. A maximum of 0.85 ± 0.07 IU/mL of filter paperase (FPase), 1.25 ± 0.04 IU/mL of carboxy methyl cellulase (CMCase) and 55.56 ± 0.52 IU/mL of xylanase activity was obtained after 7 days of incubation at 28 ± 0.5 °C using delignified S. spontaneum as carbon source under submerged fermentation conditions. Enzymatic hydrolysate of S. spontaneum was then tested for ethanol production under batch and repeated batch production system using “in-situ” entrapped Saccharomyces cerevisiae VS₃ cells in S. spontaneum stalks (1 cm × 1 cm) size. Immobilization was confirmed by the scanning electron microscopy (SEM). Batch fermentation of VS₃ free cells and immobilized cells showed ethanol production, 19.45 ± 0.55 g/L (yield, 0.410 ± 0.010 g/g) and 21.66 ± 0.62 g/L (yield, 0.434 ± 0.021 g/g), respectively. Immobilized VS₃ cells showed maximum ethanol production (22.85 ± 0.44 g/L, yield, 0.45 ± 0.04 g/g) up to 8th cycle during repeated batch fermentation followed by a gradual reduction in subsequent cycles of fermentation.     

Cool gleaners: Thermoregulation in sympatric bat species

Thermoregulatory behavior in temperate bats is influenced by gender, food availability, ambient temperature and reproduction. Ecologically and morphologically similar bat species (Myotis bechsteinii, M. nattereri, and Plecotus auritus; Vespertilionidae) facing similar diurnal conditions should therefore not differ in their thermoregulatory behavior. Identified day roosts (n = 23) of radio-tagged bats (n = 30) were spread over an area of 33.1 ha, but ambient temperature did not differ between roosting sites. Furthermore, there was no significant difference in cardinal direction, roost height, canopy coverage, and breast height diameter between day roosts used by the three species. Minimum roost temperatures and isolation values, however, differed significantly between our species with lowest values in P. auritus. The range of skin temperatures (min–max) recorded by temperature-sensitive transmitters was not species-specific with the lowest ranges in late pregnancy (mean ± SD: 7.1 ± 1.1 °C) and highest in post-lactation (mean ± SD: 13.1 ± 1.1 °C). The minimum skin temperature, however, was species-specific with the lowest values in P. auritus (mean ± SD: 20.2 ± 1.1 °C), intermediate in M. nattereri (mean ± SD: 23.4 ± 1.0 °C), and the highest in M. bechsteinii (mean ± SD: 26.8 ± 1.0 °C). Species-specific usage of energy-saving mechanisms might represent an important niche differentiation of species. Different mechanisms might allow, e.g. one species to occupy colder roosts with higher temperature variations or to shorten foraging times due to distinct thermoregulatory behavior.     

Male and female Popillia quadriguttata (Fabricius) and Protaetia brevitarsis (Lewis) (Coleoptera: Scarabaeidae) response to Japanese beetle floral and pheromone lures

Popillia quadriguttata (Fabricius), and Protaetia brevitarsis (Lewis) adults were captured with Japanese beetle, Popillia japonica Newman, sex attractant and floral lures at Changchun, China during July–August 2012. The floral lure (phenethyl propionate:eugenol:geraniol, 3:7:3) was attractive to male and female P. quadriguttata (AV: 1.2 ± 0.9; 1.1 ± 0.3; total: 2.3 ± 0.8), and was similar in attraction to the combination of the sex attractant (SA) [(R, Z)-5-(1-decenyl) dihydro-2(3H)-furanone] plus the floral lure for male (1.60 ± 0.2), female (1.30 ± 1.1) and total captures (2.9 ± 3.0). However, the SA alone captured only males in much higher numbers than when combined with the floral lure (10.0 ± 6.4). In a separate earlier test, the greatest number of P. quadriguttata males (12.5 ± 3.0), female (12.2 ± 1.5) and total captures (24.7 ± 2.5) was in yellow, laboratory-made, bottle traps. The floral lure also attracted female Pro. brevitarsis (10.0 ± 3.4), while the SA attracted only few male beetles (1.0 ± 0.2). The combination SA + floral lure captured similar females (11.0 ± 2.0) and total (14.2 ± 2.2) Pro. brevitarsis as the floral lure alone. Two butterflies, Colias erate poliographus (Motschulsky) and Pieris rapae (Linnaeus), were also attracted to the floral lure. These studies indicate a potential for replacing pesticides by using the Japanese beetle lures for monitoring and control of several insects in China, and that they would be useful in monitoring and eradication of two potential scarab pests, P. quadriguttata and Pro. brevitaris, in the United States and Europe.     

The preparation and antihyperlipidaemic assay of piperlonguminine in vivo

The natural product piperlonguminine (GBN) which is extracted from Piper longum Linn., has high antihyperlipidaemic activity and low toxicity. However, the content of natural GBN in P. longum (0.20–0.25%) is low, and it is not easy to prepare enough sample of natural GBN for further studies, such as large-scale animal experiments. Therefore, in the present study, we tested and confirmed the antihyperlipidaemic activity of chemically synthesised GBN in rats for the first time. The results of the antihyperlipidaemic assay in vivo showed that synthetic GBN had significant lipid-lowering activities. Synthetic GBN not only inhibited body weight gain (66.3 ± 22.50 g vs. 83.9 ± 19.95 g) but also significantly decreased total cholesterol (TC, 9.67 ± 3.32 mmol/L vs. 22.26 ± 5.84 mmol/L), total glycerol (TG, 1.47 ± 0.08 mmol/L vs. 2.86 ± 0.75 mmol/L), and low-density lipoprotein cholesterol (LDL-C, 3.57 ± 1.15 mmol/L vs. 5.44 ± 1.42 mmol/L) while increasing high-density lipoprotein cholesterol (HDL-C, 1.31 ± 0.56 mmol/L vs. 0.68 ± 0.20 mmol/L) in the serum of rats fed a high-fat diet.     

Differences in preovulatory follicle dynamics and timing of preovulatory LH surge affect fertility of maiden sheep reared in semi-arid extensive conditions

In the current study follicular dynamics, pituitary function, ovulatory response and luteal activity of 30 maiden Barbarine sheep were analyzed according to oestrus occurrence and lambing outcome after oestrus synchronisation with cloprostenol. Animals were retrospectively classified in three groups named as O? (n = 7, ewes not displaying oestrus), O+L? (n = 7, ewes showing oestrus but failing to lamb) and O+L+ (n = 16; ewes showing oestrus and lambing thereafter). All the sheep ovulated and daily transrectal ultrasonographies revealed that preovulatory follicles were present at cloprostenol injection in all the animals. In sheep O+L+ and O+L?, 50% and 57% of the ovulatory follicles were the largest follicles at cloprostenol treatment (mean size of 4.1 ± 0.26 mm and 4.3 ± 0.74 mm, respectively). In O? ewes, the same percentage was higher (86%, P < 0.05 when compared to group O+L+; mean size of 4.0 ± 0.46 mm). The number of large follicles and the final diameter of the ovulatory follicles at oestrous tended thereafter to be higher in group O+L+ (1.4 ± 0.1 and 6.4 ± 0.2) than in groups O+L? (1 ± 0.2 and 5.7 ± 0.36) and O? (0.9 ± 0.2 and 5.9 ± 0.5, respectively). Conversely, the number of medium follicles at oestrus detection was higher in the group O+L? (2.1 ± 0.3, P < 0.05) than in the other two groups (1 ± 0.2 and 1 ± 0.3 for O+L+ and O? respectively). Timing of preovulatory LH surge was earlier for ewes O? (24.0 ± 4.75, P < 0.05) than for sheep O+L+ and O+L? (37.9 ± 2.45 h and 38.0 ± 4.75 h, respectively) and 94% of O+L+ ewes had a LH surge between 16 h and 64 h after cloprostenol injection compared to 57% in O+L? and O? groups (P < 0.05). Thus, maiden Barbarine sheep failing to display oestrus or conceive showed alterations in their follicular dynamics and, thereafter, pituitary function and ovulatory response.     

The effect of diet on longevity,fecundity, and the sex ratio of Clitostethus arcuatus (Rossi) (Coleoptera: Coccinellidae)

Clitostethus arcuatus is a major, cosmopolitan predator of some Aleyrodidae. Field collected adult beetles were reared in the laboratory on different diets: Siphoninus phillyreae eggs, Trialeurodes vaporariorum eggs, Sitotroga cerealella eggs, or an artificial diet consisting of honey, yeast, and pollen. All experiments were conducted at 25 ± 2 °C, 65 ± 5% RH and a photoperiod of 16:8 (L:D) h. Female and male C. arcuatus consumed a mean (± SE) of 61 ± 0.6 and 27 ± 0.9 T. vaporariorum eggs d^? 1, respectively, and a mean of 56 ± 2.2 and 29 ± 1.1 S. phillyreae eggs d^? 1, respectively. Significant differences were noted between sexes and between hosts consumed by female C. arcuatus. No feeding occurred on S. cerealella eggs. Although there was a significant difference between rates of oviposition due to diet, fertility rates on different diets did not show significant differences. The sex ratio of C. arcuatus (female:male) was 51.4:48.6, 55.2:44.8, and 54.6:45:4 when adults fed on T. vaporariorum, S. phillyreae, and artificial diet, respectively. These differences were not significantly different. Average longevity (± SE) was 66.4 ± 2.6, 54.9 ± 2.5; 77.3 ± 6.9, 67.5 ± 7.2; and 86.4 ± 4.5 70.3 ± 3.6 days for female and male C. arctuatus, respectively, on T. vaporariorum, S. phillyreae and artificial diet, respectively, with significant differences between sexes and diets. Although developmental duration on T. vaporariorum was longer than ash whitefly, this difference was not significant (mean 27.68 ± 0.31 and 25.09 ± 0.21 days for predators reared on T. vaporariorum and S. phillyreae, respectively). Given its longevity and fecundity on T. vaporariorum, C. arcuatus may be a good choice for mass release on glasshouse crops infected by greenhouse whitefly.