Embryonic and larval development of garpike from the Adriatic Sea

Although information about embryonic and larval development of garpike, Belone belone (Linnaeus, 1761), is present in the published literature, the bulk of research concerns garpike from the northeastern Atlantic Ocean and the Baltic Sea. The present work describes the embryonic and larval development of garpike, Belone belone, from the Adriatic Sea, and methods used for incubation of fertilized eggs in aquarium conditions. Because garpike is, as suggested by some authors, divided into subspecies, we conclude that some differences in embryonic development could also be expected. In the present study, eggs were fertilized using the dry fertilization method and were incubated in a tank equipped with aeration and constant sea water flow. Salinity and content of dissolved oxygen were constant, and the temperature varied between 19.4 and 22.3°C. Eggs were spherical, measuring 3071.9 ± 75.73 μm in diameter. Yolk sacs were homogeneous and did not contain oil globules. The first larvae hatched 329 h and 47 min after fertilization. Absorption of the yolk sac occurred 17 h – 48 h after hatching and the total length of newly hatched larvae was 9.78 mm. The peculiarities observed in the embryonic and early larval development are evidence of an exceptional plasticity and adaptive potential, which could be considered as helpful features in extending the natural range of occurrence of this species.     

Isolation and Characterization of Potentially Human Pathogenic,Cytotoxin‐producing Aeromonas Strains from Retailed Seafood in Berlin,Germany

The presence of potentially human pathogenic strains of Aeromonas was investigated in 84 samples of seafood which were purchased from retail traders in Berlin, Germany in spring 2000. A total of 134 Aeromonas strains were isolated on selective [GSP agar and Aeromonas (Ryan) agar] and unselective (standard count agar and enterohaemolysin agar) media from 27 (32.1%) of the samples and were classified as Aeromonas hydrophila (67.9%), A. caviae (26.1%) and A. sobria (6.0%) by biotyping. Thirteen (48.1%) of the 27 positive samples contained more than one species of Aeromonas. Production of haemolysins on enterohaemolysin agar was found with 132 (98.5%) of the strains at 28°C and with 130 strains (97.0%) at 37°C growth temperature. Vero cytotoxins were produced by 99 (73.9%) of the strains when grown at 28°C but only by 24 of the strains (17.9%) at 37°C. The latter strains were identified as A. hydrophila (n = 22) and A. sobria (n = 2) which came from 17 (20.2%) samples of raw seafood and from ready‐to‐eat salted herring ‘Matjes’ products. Cytotoxin‐encoding genes for aerolysin (aer) and haemolysin A (hlyA) were investigated by PCR. Aer and hlyA genes were detected in both, strains which produced toxins only at 28°C and strains which produced toxins at 37°C. Our data indicate that raw seafood and ready‐to‐eat fish products can harbour potential human pathogenic, cytotoxin producing Aeromonas strains.     

Effects of electrical stimulation and pre‐rigor conditioning temperature on aging potential of hot‐boned beef M. longissimus lumborum

The objective of this study was to create various pH/temp decline rates in hot‐boned bull beef M. longissimus lumborum (LL) through a combination of electrical stimulation (ES) and pre‐rigor holding temperature. The relationship between the pre‐rigor interventions, the activities of µ‐calpain and small heat shock proteins (sHSP), and the impacts on meat product quality were determined. Paired LL loins from 13 bulls were hot‐boned within 40 min of slaughter, immediately ES and subjected to various holding temperatures (5, 15, 25, and 35°C) for 3 hr. The rate of muscle pH decline, sarcomere length, shear force, and proteolysis of muscle proteins were measured. ES‐25°C had a longer sarcomere length compared to non‐electrical stimulation samples. ES‐25°C and ES‐35°C samples had lower shear force values, higher µ‐calpain activity and higher desmin, troponin‐T, and sHSP degradation. The above findings suggest that pH/temp decline rates created in hot‐boned muscle impacted muscle protein proteolysis by increasing the activity of proteases and degradation of sHSP.     

Determination of sulfadiazine,trimethoprim, and N4‐acetyl‐sulfadiazine in fish muscle plus skin by Liquid Chromatography–Mass Spectrometry. Withdrawal‐time calculation after in‐feed administration in gilthead sea bream (Sparus aurata L.) fed two different diets

This study presents a depletion study for sulfadiazine and trimethoprim in muscle plus skin of gilthead sea bream (Sparus aurata L.). N⁴‐acetyl‐sulfadiazine, the main metabolite of sulfadiazine (SDZ), was also examined. The fish were held in seawater at a temperature of 24–26 °C. SDZ and trimethoprim (TMP) were administered orally with medicated feed for five consecutive days at daily doses of 25 mg SDZ and 5 mg TMP per kg of fish body weight per day. Two different diets, fish oil‐ and plant oil‐based diets, were investigated. Ten fish were sampled at each of the days 1, 3, 5, 6, 8, 9, 10, and 12 after the start of veterinary medicine administration. However for the calculation of the withdrawal periods, sampling day 1 was set as 24 h after the last dose of the treatment. Fish samples were analyzed for SDZ, TMP, and acetyl‐sulfadiazine (AcSDZ) residues by liquid chromatography–mass spectrometry. SDZ and TMP concentrations declined rapidly from muscle plus skin. Considering a maximum residue limit of 100 μg/kg for the total of sulfonamides and 50 μg/kg for TMP residues in fish muscle plus skin, the withdrawal periods of the premix trimethoprim‐sulfadiazine 50% were calculated as 5 and 6 days, at 24–26 °C, in fish oil (FO) and plant oil (PO) groups, respectively. The investigation of this work is important to protect consumers by controlling the undesirable residues in fish.     

In Vitro Evaluation of Liquid‐stored Buffalo Semen at 5°C Diluted in Soya Lecithin Based Extender (Bioxcell®), Tris‐Citric Egg Yolk,Skim Milk and Egg Yolk‐Citrate Extenders

This study was designed to compare the quality of liquid‐stored buffalo bull spermatozoa in soya lecithin based extender Bioxcell^® (BIOX), milk (MILK), tris‐citric egg yolk (TEY) and egg yolk‐citrate (EYC) extender at 5°C. Semen was collected from five Nili‐Ravi buffalo (Bubalus bubalis) bulls of 6–7 years of age with artificial vagina over a period of 3 weeks (two consecutive ejaculates once in a week). Semen ejaculates having more than 60% motility were pooled, split into four aliquots, diluted (37°C; 10 × 10⁶ motile spermatozoa/ml), cooled from 37 to 5°C in 2 h (0.275°C/min) and stored for 5 days. Sperm motility, viability, plasma membrane integrity (PMI) and normal acrosomal ridge were studied at first, third and fifth day of storage. Higher values of progressive sperm motility (%), sperm viability (%), sperm PMI (%) and normal apical ridge (%) were observed in BIOX, MILK and TEY extenders at first, third and fifth day of storage than EYC extender. Progressive sperm motility, sperm viability and sperm PMI in BIOX^® extender were not different from MILK and TEY extenders at 1st and third day storage period. However, at fifth day of storage, the values for these parameters remained significantly higher (p < 0.05) in BIOX^® compared with MILK, TEY and EYC extenders. At fifth day of storage, the semen quality parameters for Bioxcell^® were comparable to those with MILK and TEY extenders at third day of storage. In conclusion, motility, viability and PMI of buffalo bull spermatozoa remained similar in Bioxcell^®, milk and TEY extender at first and third days of storage at 5°C. Yet, the values for the aforementioned parameters in Bioxcell^® were higher compared with milk, TEY and EYC extender at fifth day of storage at 5°C.     

Protocol Optimization for Long‐Term Liquid Storage of Goat Semen in a Chemically Defined Extender

A specific problem in the preservation of goat semen has been the detrimental effect of seminal plasma on the viability of spermatozoa in extenders containing egg yolk or milk. The use of chemically defined extenders will have obvious advantages in liquid storage of buck semen. Our previous study showed that the self‐made mZAP extender performed better than commercial extenders, and maintained a sperm motility of 34% for 9 days and a fertilizing potential for successful pregnancies for 7 days. The aim of this study was to extend the viability and fertilizing potential of liquid‐stored goat spermatozoa by optimizing procedures for semen processing and storage in the mZAP extender. Semen samples collected from five goat bucks of the Lubei White and Boer breeds were diluted with the extender, cooled and stored at 5°C. Stored semen was evaluated for sperm viability parameters, every 48 h of storage. Data from three ejaculates of different bucks were analysed for each treatment. The percentage data were arcsine‐transformed before being analysed with anova and Duncan’s multiple comparison test. While cooling at the rate of 0.1–0.25°C/min did not affect sperm viability parameters, doing so at the rate of 0.6°C/min from 30 to 15°C reduced goat sperm motility and membrane integrity. Sperm motility and membrane integrity were significantly higher in semen coated with the extender containing 20% egg yolk than in non‐coated semen. Sperm motility, membrane integrity and acrosomal intactness were significantly higher when coated semen was 21‐fold diluted than when it was 11‐ or 51‐fold diluted and when extender was renewed at 48‐h intervals than when it was not renewed during storage. When goat semen coated with the egg yolk‐containing extender was 21‐fold diluted, cooled at the rate of 0.07–0.25°C/min, stored at 5°C and the extender renewed every 48 h, a sperm motility of 48% was maintained for 13 days, and an in vitro‐fertilizing potential similar to that of fresh semen was maintained for 11 days.     

Effects of Thawing Temperature and Post‐thaw Dilution on the Quality of Cat Spermatozoa

The present study aimed to compare cat sperm quality after thawing using two different temperatures (37 and 70°C) and to investigate the effects of post‐thaw dilution on the sperm quality and longevity of ejaculated cat spermatozoa. Six ejaculates of each of six male cats were collected using an electroejaculator (total 36 ejaculates). The semen was frozen in 0.25‐ml straws using a Tris egg yolk extender containing Equex STM paste. Four straws prepared from each ejaculate were thawed at four different occasions; (i) at 37°C for 15 s, (ii) at 37°C for 15 s and diluted 1 : 2 with Tris buffer (v/v), (iii) at 70°C for 6 s, (iv) at 70°C for 6 s and diluted 1 : 2 with Tris buffer (v/v). The percentages of motile spermatozoa, the scores of progressive motility, the percentages of spermatozoa with intact plasma membrane (using SYBR‐14/EthD‐1 stains) and intact acrosome (using fluorescein isothiocyanate conjugated peanut agglutinin/propidium iodide stains) were evaluated in fresh semen at 0, 2, 4 and 6 h after thawing. The thawing temperature had no effect on any sperm parameters throughout the incubation period (p > 0.05). The dilution after thawing improved sperm motility, progressive motility and acrosome integrity (p < 0.05). The thawing of cat spermatozoa and subsequently diluting with Tris buffer resulted in an immediate (at 0 h) overall (combined over temperature) percentage of motile sperm of 64.8 ± 10.7 (mean ± SD), a score of progressive motility of 4.0 ± 0.5, a percentage of spermatozoa with intact plasma membrane of 64.4 ± 12.1 and intact acrosome of 44.8 ± 20.2. In conclusion, frozen cat semen can be thawed either at 37 or 70°C and post‐thaw dilution is recommended to reduce the toxic effect of some ingredients in the extender during post‐thaw incubation.     

Pharmacokinetics of florfenicol in blunt‐snout bream (Megalobrama amblycephala) at two water temperatures with single‐dose oral administration

The pharmacokinetics and residue elimination of florfenicol (FFC) and its metabolite florfenicol amine (FFA) were studied in healthy blunt‐snout bream (Megalobrama amblycephala, 50 ± 10 g). The study was conducted with a single‐dose (25 mg/kg) oral administration at a water temperature of 18 or 28°C, while in the residue elimination study, fish were administered at 25 mg/kg daily for three consecutive days by oral gavage to determine the withdrawal period (WDT) at 28°C. The FFC and FFA levels in plasma and tissues (liver, kidneys and muscle) were analysed using high‐performance liquid chromatography (HPLC). A no‐compartment model was used to analyse the concentration versus time data of M. amblycephala. In the two groups at 18 and 28°C, the maximum plasma concentration (C_max) of FFC was 5.89 and 6.21 μg/ml, while the time to reach C_max (T_max) was 5.97 and 2.84 hr, respectively. These suggested that higher temperature absorbed more drug and more quickly at M. amblycephala. And the elimination half‐life (T_1/2kβ) of FFC was calculated as 26.75 and 16.14 hr, while the total body clearance (CL) was 0.09 and 0.15 L kg^?1 hr^?1, and the areas under the concentration–time curves (AUCs) were 265.87 and 163.31 μg hr/ml, respectively. The difference demonstrated that the elimination rate of FFC in M. amblycephala at 28°C was more quickly than that at 18°C. The results of FFA showed the same trend in tissues of M. amblycephala. After multiple oral doses (25 mg/kg daily for 3 days), the k (eliminate rate constant) of FFA in M. amblycephala muscle was 0.017, the C₀ (initial concentration) was 3.07 mg/kg, and the WDT was 10 days (water temperature 28°C).     

Effects of Temperature on In Vitro Short‐Term Storage of Sterlet Sturgeon (Acipenser Ruthenus) Ova

Artificial propagation of sturgeons is becoming increasingly important for recovery efforts as well as for commercial production. Sterlet Acipenser ruthenus is a common Eurasian sturgeon with a small body size and one of the fastest reproductive cycles among the sturgeons. The practical question being addressed in this study was how long fertilization of ovulated eggs can be delayed without substantially reducing the hatching rate, and an ancillary question is under what' temperature conditions do eggs retain good quality. Broodstock were injected with homogenized carp pituitary extract (CPE); ovulated eggs from three females were allocated to various treatment groups for temperature storage (control, 7°C, 11°C, 15°C and 19°C) until fertilized. Storage times at the regulated temperatures prior to fertilization were for 2.5, 5.0, 7.5 and 10.0 h. After the selected storage times in ovarian fluid, eggs were fertilized and transferred to incubation cages and then they were counted. Three replicates were allocated to each storage period and temperature. Hatched larvae were counted at 7‐day post‐fertilization. We found that sterlet eggs do not need to be fertilized immediately after collection. Reasonably good quality was retained for several hours if temperature conditions are fairly cool and stable. Eggs retained good quality when stored at 7°C and 11°C for up to 10 h with 54.1 ± 2.9 to 69.9 ± 7.9% hatching success, but egg quality was significantly reduced after 5‐h storage at 19°C (p < 0.01) and 7.5‐h storage at 15°C (p < 0.05) compared to cooler temperatures. Uniform temperatures between 7°C and 11°C can be considered as appropriate for storage of eggs in ovarian fluid for up to 10 h. This information can have practical application to routine hatchery practice for acipenserids, as well as for certain research protocols.     

Production of a bacteriocin‐like inhibitory substance by Leuconostoc mesenteroides subsp. dextranicum 213M0 isolated from Mongolian fermented mare milk,airag

Strain 213M0 was selected with productivity of a bacteriocin‐like inhibitory substance (BLIS) among 235 strains of lactic acid bacteria (LAB) isolated from Mongolian fermented milk ‘airag’. Strain 213M0 was species‐identified as Leuconostoc mesenteroides subsp. dextranicum by morphological observation, carbohydrate fermentation profiling and sequencing the 16S rRNA gene. Incubation temperature proper to produce the BLIS was 25°C rather than 30 and 37°C, and the production actively proceeded during the exponential growth phase of the producer cells. Antibacterial effect of BLIS 213M0 was limited to all nine strains of Listeria sp. bacteria and seven strains of LAB cocci among 53 tested strains, which corresponds to a typical feature of the class IIa pediocin‐like bacteriocins. BLIS 213M0 was not inactivated in every broad pH range solution (pH 2.0‐11.0), and was stable against storage at 25°C for 1 week and heating at 121°C for 15 min under pH 4.5. Peptide frame of BLIS 213M0 was confirmed by inactivation with some peptidases, and then its molecular weight was estimated to be 2.6‐3.0 kDa using an in situ activity assay following sodium dodecyl sulfate polyacrylamide gel electrophoresis. The estimated size was different from the other Leuconostoc bacteriocins already reported. These results suggest that BLIS 213M0 would be a novel listericidal bacteriocin.     

Effects of high ambient temperature on urea‐nitrogen recycling in lactating dairy cows

Effects of exposure to hot environment on urea metabolism were studied in lactating Holstein cows. Four cows were fed ad libitum a total mixed ration and housed in a temperature‐controlled chamber at constant moderate (18°C) or high (28°C) ambient temperatures in a cross‐over design. Urea nitrogen (N) kinetics was measured by determining urea isotopomer in urine after single injection of [¹⁵N₂]urea into the jugular vein. Both dry matter intake and milk yield were decreased under high ambient temperature. Intakes of total N and digestible N were decreased under high ambient temperature but urinary urea‐N excretion was increased. The ratio of urea‐N production to digestible N was increased, whereas the proportion of gut urea‐N entry to urea‐N production tended to be decreased under high ambient temperature. Neither return to the ornithine cycle, anabolic use nor fecal excretion of urea‐N recycled to the gut was affected by ambient temperature. Under high ambient temperature, renal clearance of plasma urea was not affected but the gut clearance was decreased. Increase of urea‐N production and reduction of gut urea‐N entry, in relative terms, were associated with increased urinary urea‐N excretion of lactating dairy cows in higher thermal environments.     

Reproduction and embryogenesis of the mandi‐amarelo catfish,Pimelodus maculatus (Pisces,Pimelodidae), in captivity

To study reproduction and embryogenesis, Pimelodus maculatus specimens were kept in captivity and captured bimonthly during 1 year. Gonads samples (211 specimens) were collected and submitted to routine histological techniques. Pimelodus maculatus prepared to reproduce when water temperature was high, and even reached advanced maturation but did not spawn in captivity. Spent fish gonads were not documented, and atretic follicles were frequent (60%) in late maturation females. When then submitted to hypophysation, 70% of the females responded positively to hormonal treatment. Oocyte extrusion occurred 8 h after a second hormonal injection at 26°C. The fertilisation rate was 65.1 ± 9.2% at 24°C. Recently spawned oocytes of P. maculatus were spherical, non‐adhesive, yellow in colour, with an average diameter of 1113.92 ± 37.02 μm and covered by a thick gelatinous layer. Blastopore closure occurred 7 h and 30 min after fertilisation. Embryonic development was completed within 18 h after fertilisation. The results of this work provide important knowledge for the handling and cultivation of not only P. maculatus, but other species of potential value for fish culture.     

Cryopreservation of Nili‐Ravi buffalo (Bubalus bubalis) semen in AndroMed® extender; in vitro and in vivo evaluation

The study was designed to evaluate AndroMed^® for the freezability and fertility of Nili‐Ravi buffalo semen. Semen was collected from four adult Nili‐Ravi buffalo (Bubalus bubalis) bulls for 3 weeks (replicate). Semen ejaculates from each buffalo bull were divided into three aliquots. One aliquot was used for evaluation of motility, plasma membrane integrity, livability, viability, DNA integrity and normal apical ridge. Remaining two aliquots were diluted (37°C; 50 × 10⁶ spermatozoa/ml) in tris‐citric egg yolk or AndroMed^® extender and cryopreserved in 0.5 ml French straws. After thawing, per cent post‐thaw motility (47.9 ± 0.8, 49.2 ± 1.7), plasma membrane integrity (44.4 ± 1.2, 46.8 ± 1.8) and normal apical ridge (81.4 ± 0.3, 83.2 ± 0.3) were recorded similar (p > .05) in tris‐citric egg yolk and AndroMed^® extender. Higher (p < .05) percentage of sperm livability (70.5 ± 1.4 and 64.4 ± 1.0), viability (67.5 ± 1.5 and 61.5 ± 0.6) and DNA integrity (97.0 ± 0.3 and 93.4 ± 0.21) were recorded in AndroMed^® compared to tris‐citric egg yolk post‐thaw. Values for all the aforementioned spermatozoal quality parameters were observed lower (p < .05) in frozen‐thawed compared to fresh semen irrespective of the experimental extenders. Fertility rates of buffalo semen did not differ (p > .05) either cryopreserved in tris‐citric egg yolk or AndroMed^® extender (45.5% vs. 49%). It is concluded that AndroMed^® is capable in protecting the buffalo bull sperm during freeze‐thawing process and can be adopted safely for routine use replacing the tris‐citric egg yolk extender in artificial insemination programme.     

IMAGING DIAGNOSIS—USE OF RADIOGRAPHY AND COMPUTED TOMOGRAPHY IN THE DIAGNOSIS OF A MINERALIZED YOLK SAC IN A BROWN KIWI (APTERYX MANTELLI)

A 12‐day‐old Brown Kiwi (Apteryx mantelli) was presented with anorexia, torticollis, head‐tilt, and coelomic distension. Radiographs showed an ill‐defined, fat‐opaque, coelomic mass displacing viscera craniodorsally. Curvilinear mineral opacities were superimposed over the ventral aspect of the mass. Computed tomography demonstrated the presence of mineral within the periphery of a fat attenuating mass consistent with a retained yolk sac. A deutectomy (yolk sac excision) was performed. Histopathology of the excised tissue confirmed the diagnosis of a retained yolk sac with multifocal mineralization.     

The Impact of Egg Ozonation on Hatching Success,Larval Growth,and Survival of Atlantic Cod,Atlantic Salmon,and Rainbow Trout

The direct exposure of fish eggs to ozonated water has generated interest as a means of ensuring pathogen-free eggs without the use of harsh chemicals. However, there are numerous knowledge gaps, including safe contact times, exposure levels, and potential long-term effects on aquaculture species in both freshwater and seawater. The effect of different ozone (O₃) doses (0.5–1.0, 1.5–2.0, and 2.5–3.0 mg of O₃/L for 90 s) on recently fertilized eggs of Atlantic Cod Gadus morhua and eyed eggs of Atlantic Salmon Salmo salar and Rainbow Trout Oncorhynchus mykiss was evaluated in comparison with the effects of two commercial disinfectants: Perosan (0.004 mg/L) and Ovadine (100 mg/L). The impact of ozone application was evaluated based on hatching success, larval nucleic acid concentration, larval growth, and survival. Overall, results indicated that ozonation of Atlantic Cod eggs at a dose less than 3.0 mg/L for 90 s produced no negative effect on the larvae up to 30 d posthatch. Furthermore, ozonation of Atlantic Salmon and Rainbow Trout eggs generated no negative effect on the larvae, based on monitoring until 85% yolk sac re-absorption (16 d posthatch).

Received May 6, 2014; accepted October 24, 2014     

Gut and intestinal passage time in the Rainbow Skink (Trachylepis margaritifer): implications for stress measures using faecal analysis

Stress levels in organisms provide a rapid measure for assessing population health. Handling and capture stress, however, cause error in blood measures, so this method is rapidly being replaced by assessing levels of stress metabolites in faeces. This eliminates the source of error because there is a lag period between stress perception and the resultant stress metabolite accumulation within faeces. This lag period is correlated with specific intestinal passage time, a measure that can vary greatly between taxa, particularly amongst ectotherms. Due to two deleterious consequences associated with extended exposure of the metabolites to the intestinal environment, species that exhibit long and variable intestinal passage times are not good candidates for metabolite studies. We measured gut and intestinal passage times in Trachylepis margaritifer to ascertain whether it would be an appropriate candidate for stress metabolite studies. We first tested if barium sulphate in the meal had an effect on gut passage time at three ambient temperatures (25, 27 and 32 °C). Barium sulphate had no effect; however, temperature had a significant effect with an unexpected pattern: gut passage time was fastest at 32 °C but was slower at 27 °C than at 25 °C. We then used X‐ray technology and barium sulphate‐loaded meals to measure gut and intestinal passage times at 25 and 27 °C. This allowed us to observe which parts of the digestive process were responsible for increased passage times at 27 °C: the faster passage time at 25 °C was due to faster intestinal passage time; there was no difference in gastric emptying time. We assess the species to be a suitable candidate for studies using faeces to measure stress. It is imperative however, that the effect of temperature on passage rates is known and taken into account in such studies.     

Characterization and partial purification of a bacteriocin‐like substance produced by thermophilic Bacillus licheniformis H1 isolated from cow manure compost

The production and partial characterization of bacteriocin‐like substances (BLSs) produced by bacteria isolated from cow manure compost were investigated. Eight BLS producers, which exhibited inhibitory activity against pathogenic bacteria, were isolated from cow manure compost at different stages of the composting process. The pile temperature ranged from 9.1°C to 73.2°C. The BLSs showed thermostability, but the BLS producers were not thermostable except for the H1 producer. Thermophilic Bacillus licheniformis H1 was further characterized. The culture supernatant of B. licheniformis H1 exhibited antagonistic activity against various species of Gram‐positive bacteria such as Listeria monocytogenes ATCC19111 but not against Gram‐negative bacteria except Pseudomonas fluorescens ATCC11251. Inactivation of bacteriocin‐like activity by α‐chymotrypsin, trypsin, and papain was highly significant (P < 0.001). The BLS was found to be stable under a pH range from 3 to 9 and at temperatures up to 75°C for 60 min, but it lost activity after being autoclaved at 121°C for 15 min. The optimum production of BLS by B. licheniformis H1 was obtained at a temperature of 55°C. Sodium dodecyl sulfate – polyacrylamide electrophoresis analysis of concentrated partially purified supernatants collected after resting the bacterial cells at 55°C revealed a bacteriocin‐like protein with a molecular mass of approximately 3.5 kDa. This study is the first report of a BLS from thermophilic B. licheniformis with an animal compost origin.     

Effects of photoperiod on salsolinol‐induced prolactin secretion in goats

The aim of the present study was to clarify the relation between salsolinol (SAL)‐induced prolactin (PRL) release and photoperiod in goats. A single intravenous (i.v.) injection of SAL was given to adult female goats under short (8 h light, 16 h dark) or long (16 h light, 8 h dark) photoperiod conditions at two different ambient temperatures (20°C or 5°C), and the PRL‐releasing response to SAL was compared to that of thyrotropin‐releasing hormone (TRH) or a dopamine (DA) receptor antagonist, sulpiride. SAL, as well as TRH or sulpiride, stimulated the release of PRL promptly after each injection in both 8‐ and 16‐h daily photoperiods at 20°C (P < 0.05). The area under the response curve (AUC) of PRL for the 60‐min period after injections of saline (controls), SAL, TRH and sulpiride in the 16‐h daily photoperiod group was greater than each corresponding value in the 8‐h daily photoperiod group (P < 0.05). There were no significant differences in the AUC of PRL among the values produced after the injection of SAL, TRH and sulpiride in 16‐h daily photoperiod group; however, the values produced after the injection of TRH were smallest among the three in the 8‐h daily photoperiod group (P < 0.05). The PRL‐releasing responses to SAL, TRH and sulpiride under a short and long photoperiod condition at 5°C resembled those at 20°C. These results show that a long photoperiod highly enhances the PRL‐releasing response to SAL as well as TRH or sulpiride in either medium or low ambient temperature in goats.     

Growth Model of a Plasmid‐Bearing Virulent Strain of Yersinia pseudotuberculosis in Raw Ground Beef

The growth kinetics of virulence plasmid‐bearing Yersinia pseudotuberculosis (YPST) in sterile ground beef were studied at temperatures ranging from 0 to 30°C. In irradiated sterile ground beef, YPST replicated from 0 to 30°C, with corresponding growth rates (GR) ranging from 0.023 to 0.622 log CFU/h at 0–25°C, and the GR was 0.236 log CFU/h at 30°C. The maximum population densities (MPD) ranged from 8.7 to 11.0 log CFU/g. The growth and MPD of YPST were reduced significantly at 30°C. Models for GR and MPD of YPST in raw ground beef (RGB) as a function of storage temperatures were produced and displayed acceptable bias and accuracy. The models were validated with rifampicin‐resistant YPST (rif‐YPST) in sterile ground beef stored at 4, 10 and 25°C. The observed GR and MPD were within 95% of the predicted values. When compared to non‐sterile retail ground beef, the growth of rif‐YPST was not inhibited and displayed similar GR at 0, 10 and 25°C and MPDs as sterile ground beef at 10 and 25°C. Moreover, there was no loss of virulence plasmid in YPST during its growth in ground beef indicating that RGB contaminated with virulence plasmid‐bearing YPST could cause disease due to refrigeration failure, temperature (10–25°C) abuse, and if the meat was not properly cooked.