Effects of Water Temperature, Density, and Sex Ratio on the Spawning Rate of Red Claw Crayfish Cherax quadricarinatus (von Martens)

Effects of water temperature on red claw crayfish Cherax quadricarinatus spawning rates was evaluated for 3 mo at 22, 26 and 30 ± 1 C. A 20/m² stocking density and a 1:2 M: F sex ratio was used. Spawning rate increased as temperature increased, with 30 C providing the highest spawning rate ( P < 0.05). Effects of stocking density and sex ratio on the reproduction of red claw crayfish were also evaluated at densities of 10.15 and 20/m², with a sex ratio of 1:1, and at three sex ratios (1:1. 1:3 and 1:5 M: F) at a density 10/m². Water temperature was maintained at 28 ± 1 C and photoperiod was maintained at 14L: 10D for 6 mo during the density and sex ratio experiments. Spawning rates were not significantly different ( P > 0.05) within the three stocking densities at a sex ratio of 1:1, nor were they different ( P > 0.05) for the three sex ratios at a density of 10/m². Results revenled that densities as high as 20/m² and sex ratios of 1 male to 5 females can be used to increase spawning per tank. Also, high temperature (28 ± 1 C) and long day lengths (14L: 10D) can be utilized to stimulate peak spawning rates, but high spawning rates cannot be maintained for more than about 3 mo.     

Broad-Spectrum Resistance to Different Geographic Strains of Papaya ringspot virus in Coat Protein Gene Transgenic Papaya

ABSTRACT Papaya ringspot virus (PRSV) is a major limiting factor for cultivation of papaya (Carica papaya) in tropical and subtropical areas throughout the world. Although the coat protein (CP) gene of PRSV has been transferred into papaya by particle bombardment and transgenic lines with high resistance to Hawaii strains have been obtained, they are susceptible to PRSV isolates outside of Hawaii. This strain-specific resistance limits the application of the transgenic lines in other areas of the world. In this investigation, the CP gene of a local strain isolated from Taiwan, designated PRSV YK, was transferred into papaya via Agrobacterium-mediated transformation. A total of 45 putative transgenic lines were obtained and the presence of the transgene in papaya was confirmed by polymerase chain reaction amplification. When the plants of transgenic lines were challenged with PRSV YK by mechanical inoculation, they showed different levels of resistance ranging from delay of symptom development to complete immunity. Molecular analysis of nine selected lines that exhibited different levels of resistance revealed that the expression level of the transgene is negatively correlated with the degree of resistance, suggesting that the resistance is manifested by a RNA-mediated mechanism. The segregation analysis showed that the transgene in the immune line 18-0-9 has an inheritance of two dominant loci and the other four highly resistant lines have a single dominant locus. Seven selected lines were tested further for resistance to three PRSV heterologous strains that originated in Hawaii, Thailand, and Mexico. Six of the seven lines showed varying degrees of resistance to the heterologous strains, and one line, 19-0-1, was immune not only to the homologous YK strain but also to the three heterologous strains. Thus, these CP-transgenic papaya lines with broad-spectrum resistance have great potential for use in Taiwan and other geographic areas to control PRSV.     

花生褐斑病菌培养基筛选及生理性状和致病性研究

 花生褐斑病菌(Cercospora arachidicola Hori)在花生秆培养基上的产孢量优于前人推荐的3种培养基。在6种供试培养基中,菌丝生长量则以前人推荐的花生叶斑病尾孢菌培养基最大。培养过程中,不同光照处理对病菌生长和产孢不敏感;不同温度处理产孢量有显著差异。2%葡萄糖液有促进孢子萌发作用。病菌经人工培养不失其致病性。     

The role of T cells in protection by an inactivated infectious bursal disease virus vaccine

The current belief is that the humoral immune response plays the principal role in defense against virulent infectious bursal disease virus (IBDV). In this study we used a model, in which chickens were compromised in functional T cells by neonatal thymectomy and Cyclosporin A (TxCsA) treatment, to demonstrate the role of T cells in protective immunity against IBDV. We demonstrated that T cells were necessary to achieve full protection against virulent IBDV. When T cell compromised TxCsA-treated chickens were vaccinated with an inactivated IBDV (iIBDV) vaccine, 91% were not protected against IBDV challenge in comparison to T cell-intact chickens, which had a protection rate of 91%. The iIBDV vaccine induced virus neutralizing (VN) and ELISA antibodies, respectively, in 65 and 5% of TxCsA-treated, and in 100 and 58% of T cell-intact birds. These observations provide evidence that the stimulation of T helper cells is needed for the production of protective antibody levels in iIBDV-vaccinated chickens. Passive administration of VN anti-IBDV antibodies inducing a circulating antibody level of log(2)8 in chickens revealed that the levels of antibodies that protected T cell-intact chickens against virulent IBDV challenge were not protective for TxCsA chickens. These results indicated that antibody alone was not adequate in inducing protection against IBDV in chickens and that T cell-involvement was critical for protection. We propose that the inability of iIBDV to protect TxCsA chickens was due to compromised T cell immunity, functional T helper cells and most likely also cytotoxic T cells are needed in iIBDV vaccine protection.     

Specific detection of<Emphasis Type="Italic">Pythium aphanidermatum</Emphasis> from hydroponic nutrient solution by booster PCR with DNA primers developed from mitochondrial DNA

Pythium aphanidermatum causes damping-off and root rot of vegetable crops in hydroponic systems. A DNA probe was isolated and modified from a library ofHindIII-digested mitochondrial DNA ofP. aphanidermatum that strongly hybridized to DNA ofP. aphanidermatum and weakly hybridized to DNA ofPythium deliense. Cross-hybridizing sequences were absent from DNA of plants and other related fungi. The probe detected as little as 5 ng ofP. aphanidermatum DNA and 250 ng ofP. deliense DNA in slot-blot assays.P. aphanidermatum was detected by a hybridization assay of total DNA extracted directly from infected roots. A pair of oligonucleotide primers P1 and RP2, which allowed amplification of a specific 0.65 kb DNA fragment ofP. aphanidermatum using polymerase chain reaction (PCR), was designed from a specific DNA probe. Specific amplification of this fragment fromP. aphanidermatum was highly sensitive, detecting template DNA as low as 0.1 pg total DNA by booster PCR. Specific booster PCR amplification using P1 and RP2 was successful in detectingP. aphanidermatum in naturally infected nutrient solution and roots of vegetables in a field hydroponic system. http://www.phytoparasitica.org posting Sept. 22, 2002.     

Influences of Follicular Size on Parthenogenetic Activation and in Vitro Heat Shock on the Cytoskeleton in Cattle Oocytes

The availability of cow ovaries from the slaughterhouse has been very limited in Taiwan. To maximize the use of cow ovaries for research purposes, whole ovary dissection was performed and the developmental competence of the oocytes derived from different sizes of follicles was assessed by the rates of in vitro maturation (IVM) and parthenogenetic activation of the oocytes in Experiment 1 (Exp 1). Cumulus-oocyte complexes (COCs) derived from small (1-2 mm) and large (3-8 mm) follicles were subjected to standard IVM culture for 24 h. Mature oocytes were selected and then parthenogenetically activated using A23187 (5 microm, 5 min) or thimerosal (200 microm, 10 min) alone or combined with 6-dimethylaminopurine (2.5 mm and 3.5 h, respectively). Activation rates of the oocytes, neither from the large nor small follicles, were affected by different activation treatments (single or combined stimuli). Whereas maturation rates for the oocytes from large follicles were superior to those from small follicles in both the single (59% vs 45%) and combined treatments (76% vs 40%; p < 0.05). To understand how prolonged heat shock (HS) influences cytoskeletal configurations of mature bovine oocytes, in Experiment 2 (Exp 2), matured oocytes derived from large follicles were randomly allocated to different durations of HS treatments at 41.5 degrees C for 0 (C0h, control, n = 12), 1 (HS1h, n = 28), 2 (HS2h, n = 31), and 4 h (HS4h, n = 30). An additional control group was cultured for 4 h without HS (38.5 degrees C, 4 h, n = 35). Alterations in nuclear structures, microtubules (MTs), and microfilaments (MFs) of the oocytes were examined. Abnormalities in the chromosomes, spindle MTs and the percentages of oocytes with cytoplasmic MTs increased with time of HS treatment. The intensity of the MF distribution in the HS oocytes was also altered. Significant changes in the cytoskeleton after HS may be associated with the reduced development under hyperthermia and, perhaps, with the low pregnancy rates of the animals during hot seasons.     

Indoor Spawning and Egg Development of the Red Claw Crayfish Cherax quadricarinatus

In 1991 and 1992, indoor hatchery experiments were conducted with red claw crayfish Cherax quadricarinatus a large tropical crayfish from northern Australia. Six fiberglass tanks were stocked with 40–190 g mature red claw crayfish at 10/m² and a 1:2 male to female ratio. Water temperature was maintained at 28 ± 1 C. Photoperiod was provided by natural daylight in 1991 and controlled lighting in 1992.
With natural lighting, monthly spawning rates (% of females spawning/month) ranged from an average of 15% during months with less than 12 h of light per day to over 35% in months with more than 12 h of light per day. Peak spawning occurred from May to July, too late to obtain juveniles for spring stocking. During the second year, photoperiod was increased from 10L:14D in December to 14L:10D in February and maintained at this level until July. Red claw spawning increased with increasing day length. Peak spawning rates of 50% per month and higher occurred from March to May. Spawning rates decreased dramatically after May, even though photoperiod and temperature remained constant. Eggs per spawn increased with an increase in female size. Fecundity average 7.3 juveniles per gram of body weight for females from 40 to 190 g, regardless of size. Egg color was useful for predicting stage of development.     

Culicoides arakawae (Diptera: Ceratopogonidae) population succession in relation to leucocytozoonosis prevalence on a chicken farm in Taiwan

Leucocytozoonosis caused by Leucocytozoon caulleryi is a significant disease prevalent in open chicken houses of southern and eastern Asia. L. caulleryi is transmitted by Culicoides arakawae, a blood-sucking vector. Leucocytozoonosis prevalence is influenced by vector population succession. Thus this examination was performed on a farm to investigate vector population succession and leucocytozoonosis prevalence in experimental chicks and to obtain the ecology data for assessing the prevalence. The findings were as follows: (1) C. arakawae adults might be highly host specific because they were rarely discovered on cattle or pig farms, and none of the experimental chickens were infected by L. caulleryi on those farms. (2) Identifying and counting gorged and gravid C. arakawae to assess leucocytozoonosis prevalence is a practical strategy. The critical vector index should be 5.0 calculated by dividing the smallest vector mean from the prevalent period by the largest vector mean from the population not causing leucotocytozoonosis. (3) Taking vector means from three or more collections each month, should be the best assessment of leucocytozoonosis prevalence because C. arakawae succession appears to have a 3-week periodicity. Hopefully, these findings will contribute to assessing leucocytozoonosis prevalence.     

Factors associated with colostrum quality in individual cows from dairy herds in the Waikato region of New Zealand

AIMS: To examine associations between various cow-level factors and quality of first-milking colostrum (measured as Brix), and to evaluate herd-level associations between vaccination against calf diarrhoea and colostrum quality, in cows from dairy herds in the Waikato region of New Zealand.

METHODS: A single colostrum sample was collected, by complete udder evacuation, from each of 20 cows from 29 dairy herds in the Waikato region of New Zealand during the 2016 spring calving period. Vaccination pre-partum with a calf diarrhoea vaccine was used in 15 herds. Each colostrum sample was tested using a digital Brix refractometer. The body condition score of each cow was recorded at the time of sample collection and farmers provided records of clinical mastitis and facial eczema from the previous 12 months, as well as the age and breed of cows. Associations between cow-level variables in non-vaccinated herds and Brix were examined using a multivariable linear mixed model and estimated marginal means obtained for different categories.

RESULTS: Mean Brix of 281 samples from cows in non-vaccinated herds was 18.7 (SD 0.26)%; 63/281 (22.4%) samples had Brix ≥22% and 152/281 (54.1%) had Brix ≥18%. Mean Brix of colostrum samples from cows aged ≥6 years (20.2 (95% CI=19.1–21.2)%) was higher than for samples from 2-year-old cows (18.6 (95% CI=17.3–19.9)%) (p=0.005). Colostrum that was collected at the first milking on the day of calving had higher Brix (20.0 (95% CI=19.1–20.9)%) than colostrum collected from cows that calved the previous day (17.5 (95% CI=16.5–18.4)%) (p<0.001). Mean Brix of colostrum samples from cows which produced ≥8?L (18.2 (95% CI=17.1–19.2)%) tended to be lower than from cows which produced <8?L first-milking colostrum (19.1 (95% CI=18.3–20.0)%) (p=0.08). Among vaccinating herds, 9/15 (60%) had ≥60% colostrum samples with Brix ≥18% compared with 4/14 (29%) of non-vaccinating herds (p=0.04).

CONCLUSIONS AND CLINICAL RELEVANCE: Colostrum quality, as measured by Brix, was associated with the total volume of first-milking colostrum, interval from calving to colostrum collection and cow age. Vaccination against calf diarrhoea was associated with a higher proportion of colostrum samples with adequate Brix. Careful selection of colostrum donor cows should ensure newborn calves are fed adequate quality colostrum which should be beneficial in preventing failure of passive transfer of IgG. Testing of colostrum from individual cows with a Brix refractometer is advocated for the selection of colostrum for feeding newborn calves.