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The effects of altering the pH of seminal fluid on the sex ratio of rabbit offspring.

It has been suggested that the pH of the vagina at the time of fertilization may have a differential effect on X- or Y-bearing sperm and thereby affect the sex of the offspring. To test this postulate, rabbit semen was collected, diluted 1:10 with a buffer of pH 5.4, 6.9, or 9.6, and after 20 minutes 0.5 ml of semen-buffer mixture was used for insemination in an ovulation-induced female. Newborn pups were examined both externally and internally for gender. The females inseminated with acidic semen had 6 litters, 50 offspring, with 48% males; those with neutral semen had 8 litters, 48 offspring, with 63% males; and those with alkaline semen had 7 litters, 49 offspring, with 49% males. There was no significant difference in these sex ratios from the expected 50% males. Motility of rabbit sperm at 23 degrees C in buffers of pH 4.6, 5.4, 6.9, 9.6, and 9.8 was reduced in vitro as the pH deviated from neutrality. Acid conditions were more detrimental than alkaline conditions. Sperm lost their motility more quickly in buffers of 37 degrees C than in buffers of 23 degrees C. It was not possible with scanning electron microscopy to distinguish morphologically between X- and Y-bearing sperm. It seems unlikely that a direct effect of pH on sperm can be a single influence on the sex of offspring.

Animals

Epizootic and enzootic aspects of Minchinia nelsoni (Haplosporida) disease in Maryland oysters.

Minichinia nelsoni disease in oysters (Crassostrea virginica) from Marumsco Bar, Pocomoke Sound, Maryland (an estuarine tributary of Chesapeake Bay) was studied for 8 years (1961-68) to determine epizootiologic relationships concerning life cycle of the parasite, pathologic effects on the host, and effects of physical factors on population density and recruitment of the host and parasite. The study period covered pre-epizootic, epizootic, and post-epizootic disease conditions. Data on the native oyster population as well as annual introductions of previously unexposed, susceptible populations of juvenile oysters from 1965-68 were included. Salinity, water temperature, mortality, prevalence, incidence, life cycle stages, gross pathology, and histopathologic relationships were observed. Mortality was high (45-55% per year) during the first 3 years of the study; however, M. nelsoni prevalences were low (less than 25%) and did not clearly imply a cause and effect relationship. Drought conditions that began in the summer of 1963 and continued through 1967 caused higher salinities, and apparently initiated epizootic disease in the native oyster populations. The epizootic peaked in May 1965 with a diagnosed prevalence in native oysters of 70%. Enzootic levels of annual mortality (40% in 1966, 30% in 1967, and 2% in 1968) and fall prevalence (16%, 24%, and 4%) developed after that time. Introduced populations had a typical epizootiologic pattern in 1965 55% annual mortality, 82% incidence) and 1966 (55% annual mortality, 66% incidence) which declined in 1967 (30% annual mortality, 44% incidence) followed by a disappearance of the disease in 1968. Epizootiologic differences noted between native oysters (adult and juvenile) and the introduced juvenile populations were also evident from the stages of the disease. Infections in native animals tended to be less serious, and in many cases were delayed or attentuated, while infections in introduced oysters progressed to advanced or terminal phases. Occult manifestations (mantle recession thought to be due to M. nelsoni in oysters not showing histologic evidence of infection) were absent in introduced populations and common in the native population. These differences are interpreted as evidence of resistance in surviving native oysters and their progency, and may indicate genetic resistance developed by natural selection and manifested by an increased ability to survive and overcome infection.

Animal Diseases

Aphid symbiotic virus engineered for in vivo expression of insecticidal effectors.

Microbial pesticides are eco-friendly alternatives to chemical pesticides. However, few viral pesticides have been developed. Insects harbor diverse symbiotic viruses, which have the potential to be engineered for translational applications in pest control. Here, we engineered Acyrthosiphon pisum virus (APV), a symbiotic virus of the pea aphid, to deliver anti-aphid effectors using reverse genetics technology. A cytomegalovirus (CMV) promoter-driven APV infectious clone was successfully rescued in pea aphids with the assistance of nanocarrier star polymer (SPc). Based on this infectious clone, the protein coding sequence of chymotrypsin inhibitor variant 8 (Chy8) and the double-stranded RNA sequence targeting the aphid clip-domain serine protease (SPLP) were separately assembled into the APV genome to generate APV-Chy8 and APV-dsSPLP infectious clones, respectively. The recombinant APV clones reduced aphid relative survival rates by 34% and 17% by microinjection, respectively. To enhance the transcriptional efficiency, the APV-Chy8 and APV-dsSPLP clones were transcribed in vitro using the T7 promoter. The in vitro-synthesized APV-Chy8 and APV-dsSPLP clones reduced aphid relative survival rates by 48% and 45% by microinjection, respectively. These results demonstrate that engineered APV can deliver cargos and reduce aphid survival under injection-based experimental conditions, highlighting the potential of symbiotic virus-based vectors for delivering insecticidal effectors.

Animals

Engineering the efficient Exo-Cas12i2 for MITE manipulation in rice.

Exo-Cas12i2 v1, a fusion of the 5' exonucleases T5E and PapE, facilitates editing of TA-rich regions and mediates deletions of large genomic fragments. Exo-Cas12i2 v1-driven MITE manipulation enables precise regulation of genes involved in gibberellin-mediated cell elongation and root ethylene responses to generate favorable agronomic traits.

Oryza

Genetic manipulation in higher organisms. I. Goldfish ova as materials of operation, mRNA mediated alteration of the liver specific isozymes.

Both crucian and carp egg-mRNAs were found to mediate the transformation of goldfish tail from the double (veil-shaped) to the single (fork-shaped). Livers of these single tailed goldfish were tested for genetic change using lactate dehydrogenase isozymes (LDH) as marker. Electrophoresis of liver extracts on starch gel showed that the liver specific LDH-C4 isozymes of both the egg-mRNA and liver-mRNA injected goldfish resembled the control goldfish except an additional intermediate band. The migration distance of this new band matched the third band of the hybrids (carp X goldfish). These findings lead to the conclusion that carp egg-mRNA contains a fraction partly responsible for liver development. Rat liver was found possessing the cathodally migrating C4 isozymes. Injection of rat liver-mRNA into goldfish eggs resulted in the intensification of the liver specific C4 isozymes. It appears therefore that the input informational molecules (liver-mRNA) and the output molecular products (liver C4 isozymes) are causally related. This causal relationship would pave the way for genetic manipulation in higher organisms.

Animals

Behavior and single gene substitution in Drosophila melanogaster. I. Mating and courtship differences with w, cn, and bw loci.

The effect of single allele substitutions into an isogenic background in Oregon-R inbred lines of Drosophila melanogaster on courtship and mating patterns has been studied. A comparison has been made between the white locus w, wco, we, the wild type w+, cn, bw, and cn bw to test the effect of eye pigmentation in influencing courtship and mating patterns. It was found that w, we, wco, cn, and bw females were more successful in mating than were wild-type and cn bw females, cn bw females being less successful than wild-type females. Also, w and cn bw males were equally successful in mating but less successful than wild-type males during the 20-min test period. The mutant males performed as well as the wild-type after courtship was initiated. The behavioral parameters measured were (1) courtship latency, the time from exposure of male to female until orientation; (2) mating speed, the time from beginning of orientation of male to female until successful copulation, and (3) copulation time.

Animals

[Present problems of somatic cell genetics].

A review of recent advances in genetics of somatic cells is given in the article. It contains three sections: 1) hybridization of somatic cells and its application to the mapping of genes and to the study of gene action; 2) relation of somatic cell genetics to genetic engineerings; 3) progress and perspectives of higher plant somatic cell genetics.

Animals

A Novel Approach to Engineering Tomato Spotted Wilt Virus Infectious Clones by Disarming Key Nodes in Antiviral Defenses.

Tomato spotted wilt virus (TSWV) is an economically devastating pathogen that rapidly overcomes genetic resistance in major crops. Reverse genetic systems are crucial for investigating plant-virus interactions and resistance-breaking mechanisms, and developing these tools for segmented ambisense RNA viruses remains a crucial challenge. Current TSWV clones rely on extensively modified Asian isolates requiring co-delivery of multiple replication helpers and viral silencing suppressors. Streamlining these systems for regionally significant strains with minimal genetic alterations is essential. Here, we developed the first infectious clone of a U.S. TSWV isolate (PA01). Three binary plasmids contain cDNAs for the antigenomic L and S segments, as well as the genomic M segment, with enhanced GFP replacing NSs on the S segment. Co-delivery of the cucumovirus 2b alone or in combination with tombusvirus P19 or begomovirus AL2 achieved a high proportion of systemically infected Nicotiana benthamiana and Capsicum annuum plants. In N. tabacum, co-delivering the Caenorhabditis elegans cell death suppressor CED-9 or using NahG transgenic plants produced 30 to 33% systemically infected plants. Co-delivery of 2b boosted infection levels in NahG plants to 62%. These data indicate that in addition to the antiviral RNA-silencing machinery, additional host defense pathways influence TSWV rescue and systemic infection from cDNA. [Formula: see text] Copyright © 2026 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Tospovirus

Semen separation technique monitored with greater accuracy by B-body test.

A fluorescent staining technique, the B-body test, is utilized to ascertain the proportion of male and female spermatozoa in separated semen. This test is also used to monitor progress of Thermal Convection Counterstreaming Sedimentation and Forced Convection Galvanization processes for separating heavier electropositive (female) and lighter electronegative (male) spermatozoa into the two constituent fractions. Biological field tests using separated semen with 1,115 cows resulted in 510 progeny showing close correlation in the percentage of male versus female and B-body positive in the male spermatozoa fractions, while a control group with nonseparated semen, usually 46% B-body positive, yielded 55% male offspring and 72.0% pregnancy. Thus, the accuracy of the separation techniques has been established to the extent of producing more females with heavier and electropositive and more males with the lighter and electronegative spermatozoa fractions. However, the pregnancy rate slopes downward as the positive electropotential and density of the spermatozoa are successively increased.

Animals