The dietary ionic effects on sex ratios in animal models

Linge, Augustine Peter Kavoo (2016-03)

Thesis (PhD)--Stellenbosch University, 2016.

Thesis

ENGLISH ABSTRACT: X-linked disorders are more expressed in male offspring and prevention of these hereditary diseases is the only recourse to date. Influencing conception towards female offspring can circumvent this problem; however sex ratio adjustment remains highly contentious. Treatment of genetic disorders through sex ratio adjustments has been examined and adopted as acceptable, easier, cheaper, safer and legal. Historically, society has been rife with allegations that diet does influence sex ratios though this has not been proven fully. The dietary chemical compositions have been claimed to act as modulators that affect the electrical charges or potential of the membranes of the oocytes and cause allosteric modification or electrotactism through a process referred to as galvanotropism and cause selective attraction towards either of the male gametes and subsequently influence the gender of the conceptus. This study was performed in an attempt to address the various questions, allegations and speculations that have been rife in many societies concerning interplay between diet, fertilization and sex ratios so as to verify the validity of these social claims by taking them to the laboratory for experimental verification. Swiss Webster mice study In a double-blind fashion, nine double groups were set up comprising of 144 families of Swiss (Webster) mice, each receiving different ionic formulations in their drinking water: 1) water and 2) glucose as controls; high serum concentrations of single elements of 3) sodium, 4) potassium, 5) calcium and 6) magnesium; combined double elements 7) sodium + potassium and 8) calcium + magnesium; and finally a cocktail of the four elements 9) sodium + potassium + calcium + magnesium. Tests included the perinatal mortality rate; the relationship between high chemical composition diet and serum levels; the effects of the study chemicals on weight gains of the study models; the effects of birth order on sex ratios and the effects of seasonal variations on sex ratios. There were 1528 deliveries with 13,040 (6,348 females and 6,692 males) pups at 8.5 pups on average per litter. a) Glucose, sodium, potassium and sodium + potassium supplementation influenced the sex ratios towards male progeny (p<0.001). Calcium (p<0.014), magnesium (p<0.008) and calcium + magnesium (p<0.001) supplementation influenced sex ratios towards the female progeny. The water (p>0.61) and cocktail solutions (p>0.0609) had no influence. b) The perinatal mortality rate was 32/1000 and was female biased among the magnesium (p<0.005) and combined calcium + magnesium (p<0.044) groups only. c) Normal serum levels were observed in the control groups (p>0.165), while significant elevated serum levels were observed among the experimental groups (p<0.0001). d) The total mean weight gains were 11.12g and 10.55g among the females and males respectively. The weight trends were used to track the general wellbeing of the animal models. e) The mean litter size was 8.5 per delivery in all the groups and generations, while no influence due to birth order were detected. f) Seasons affect the litter size, in particular the rainy season, but not the gender ratios (p>0.061). Cat fish study Parallel double blind studies looking at the dietary chemical ionic effects on the oocyte membrane electrical potential were done utilising a cat fish model (n=108). The study sought to find out effects of the following solutions on the oocyte electrical charges: 1) plain electrolyte solution, 2) glucose solution 3) sodium solution 4) potassium solution 5) calcium solution 6) magnesium solution 7) sodium + potassium solution 8) calcium + magnesium solution and the 9) cocktail solution of the four elements combined. The results revealed that oocytes retrieved from the two control groups had baseline oval polar attraction significantly more towards the positive than the negative pole (p<0.0003). There was however more significant oocyte polar attraction towards the positive electrode among the oocytes retrieved from the sodium, potassium and the combined sodium + potassium solutions (p<0.0001). Oocytes retrieved from calcium, magnesium and combined calcium + magnesium solutions had significant affinity towards the negative electrode and minimal affinity towards the positive pole (p<0.0001). Oocytes harvested from the solutions constituted with all the salts demonstrated dual attraction with more attraction to the positive electrode than the negative electrode but of no statistical significance (p>0.0530). These study findings do confirm the social allegations that a positive relationship does exist between dietary components and sex ratios. The chemicals acted as the dietary modulators that ultimately influenced the electrical cellular gametal charges and subsequently the resulting progeny. Our platform of comfort is unlike artificial sex ratio adjustment methods; the natural sex ratio adjustment methods that include the dietary method under scrutiny in this study are practiced always at the comfort of many people’s homes and are difficult to quantify or have legislation on. However, this study shows that their long term effects conform to the Fishers principle of evolution towards 1:1 sex ratios and therefore do not have significant social gender skewing on a long term basis. The study clearly explains the molecular basis upon which ions of single valency attracts the Y-bearing sperm leading to a male conceptus and how cations of double valency attracts the X-bearing sperm leading to a female conceptus despite being positively charged. The study further reaffirms the natural feminine supremacy by demonstrating that it is the ova and by extension the woman who determines the sex of the conceptus. The study ultimately confirms that the dietary ionic effects on sex ratios can be used for prevention of X-linked disorders.

AFRIKAANSE OPSOMMING: X-gekoppelde afwykings is meer uitgesproke in die manlike nageslag en voorkoming van hierdie oorerflike siektes is die enigste behandelingsopsie tot op datum. Deur bevrugting te beïnvloed om aanleiding te gee tot ‘n vroulike nageslag kan hierdie probleem omseil; maar geslagsverhouding aanpassing bly hoogs omstrede. Behandeling van genetiese afwykings deur geslagsverhouding aanpassings was ondersoek en aangeneem as aanvaarbaar, makliker, goedkoper, veiliger en wettig. Histories was die samelewing deurspek met bewerings dat dieet geslagsverhoudings kan beïnvloed, alhoewel dit nog nie ten volle bewys kon word nie. Daar was beweer dat die chemiese samestelling van die dieet as modulators kan optree wat die elektriese ladings of potensiaal van die oösiete se membrane kan affekteer; dit veroorsaak allosteriese modifikasie of elektrotaktisme deur ʼn proses waarna verwys word as galvanotropisme en veroorsaak selektiewe aantrekking van twee verskillende tipes manlike gamete wat dus die geslag van die embrio beïnvloed. Hierdie studie was uitgevoer in ʼn poging om die verskillende vrae, bewerings en bespiegelings aan te spreek wat algemeen in verskeie gemeenskappe voorkom met betrekking tot die interaksie tussen dieet, bevrugting en geslagsverhoudings ten einde die geldigheid van hierdie sosiale aansprake te verifieer deur dit in die laboratorium na te vors vir eksperimentele verifikasie. Switserse Webster muis-studie Tydens ʼn dubbel-blinde studie is nege groepe bestaande uit 144 gesinne van Switserse (Webster) muise opgestel; elke groep het verskillende ioniese formulerings in hulle drinkwater ontvang: 1) water en 2) glukose as kontroles; hoë serum konsentrasies van enkele elemente van 3) natrium, 4) kalium, 5) kalsium en 6) magnesium; gekombineerde dubbele elemente, 7) natrium + kalium en 8) kalsium + magnesium; en uiteindelik 'n mengsel van al vier die elemente 9) natrium + kalium + kalsium + magnesium. Toetse het ingesluit die bepaling van die perinatale sterftesyfer; die verhouding tussen ʼn hoë chemiese samestelling dieet en serum vlakke; die gevolge van die bestudeerde chemikalieë op die gewigstoename van die studie modelle; die gevolge van geboorte-orde op geslagsverhoudings en die gevolge van seisoenale variasies op geslagsverhoudings. Daar was 1528 geboortes met 13040 (6348 vroulik en 6692 manlik) kleintjies met gemiddeld 8.5 kleintjies per werpsel. a) Glukose, natrium, kalium en natrium + kalium aanvullings het die geslagsverhoudings oorwegend na manlike nageslag beïnvloed (p<0.001). Kalsium (p<0.014), magnesium (p<0.008) en kalsium + magnesium (p<0.001) aanvullings het die geslagsverhoudings oorwegend na vroulike nageslag beïnvloed. Die water (p>0.61) en die gekombineerde mengsel oplossing (p>0.0609) het geen invloed getoon nie. b) Die perinatale sterftesyfer was 32/1000 en was vroulik bevooroordeeld onder die magnesium (p<0.005) en gekombineerde kalsium + magnesium (p<0.044) groepe alleenlik. c) Normale serum vlakke was waargeneem in die kontrole groepe (p>0.165), terwyl aansienlike verhoogde serum vlakke onder die eksperimentele groepe (p<0.0001) waargeneem was. d) Die totale gemiddelde gewig aanwins was 11.12g en 10.55g onder onderskeidelik die vroulike en manlike muise. Die gewig tendense is gebruik om die algemene welsyn van die dier-modelle te monitor. e) Die gemiddelde werpselgrootte was 8.5 per geboorte vir al die groepe en generasies, terwyl geboorte-orde geen invloed getoon het nie. f) Seisoene beïnvloed die werpselgrootte, in besonder die reënseisoen, maar nie die geslagsverhoudings nie (p>0.061). Katvis studie Parallelle dubbel-blinde studies is gebruik om die chemiese ioniese uitwerking van die dieet op die oösiet membraan se elektriese potensiaal te bepaal deur gebruik te maak van ‘n katvis model (n=108). Die doel van die studie was om die uitwerking van die volgende oplossings op die oösiet se elektriese ladings te bepaal: 1) gewone elektroliet oplossing, 2) glukose oplossing 3) natrium oplossing 4) kalium oplossing 5) kalsium oplossing 6) magnesium oplossing 7) natrium + kalium oplossing 8) kalsium + magnesium oplossing en die 9) mengsel van die vier elemente gekombineer. Die resultate het aangetoon dat oösiete wat van die twee kontrolegroepe ingewin is basislyn ovaalmembraan polêre aantrekkingskrag het wat betekenisvol meer neig na die positiewe as die negatiewe pool (p<0.0003). Daar was egter meer betekenisvolle polêre oösiet aantrekkingskragte na die positiewe elektrode onder die oösiete wat ingewin is vanuit die natrium, kalium en die gekombineerde natrium + kalium oplossingsgroepe (p<0.0001). Oösiete ingewin vanuit die kalsium, magnesium en gekombineerde kalsium + magnesium oplossingsgroepe het ʼn beduidende verhoogde affiniteit teenoor die negatiewe elektrode getoon en minimale affiniteit teenoor die positiewe pool (p<0.0001). Oösiete wat geoes was vanuit die oplossingsgroepe wat saamgestel was met al die soute het ʼn tweeledige aantrekkingskrag gedemonstreer met meer aantrekking na die positiewe elektrode as die negatiewe elektrode, maar was van geen statistiese belang nie (p>0.0530). Die bevindinge in hierdie studie bevestig die sosiale bewerings dat daar wel ʼn positiewe verhouding bestaan tussen dieet komponente en geslagsverhoudings. Die chemikalieë het opgetree as die dieet modulators wat uiteindelik ʼn invloed gehad het op die elektriese sellulêre gameet ladings en die gevolglike nageslag. Ons platform van gerief is in teenstelling met kunsmatige geslagsverhouding aanpassing metodes; die natuurlike geslagsverhouding aanpassing metodes wat die dieet metode in hierdie studie onder die soeklig plaas, word altyd uitgevoer in die gerief van talle mense se huise en is moeilik om te kwantifiseer of wetgewing oor te hê. Hierdie studie toon dat hulle langtermyn-effekte ooreenstem met die Fischer beginsel van evolusie wat neig na ʼn 1:1 geslagverhouding en dus nie 'n beduidende sosiale geslags-skeefheid op 'n langtermyn basis toon nie. Die studie omskryf duidelik die molekulêre basis waarop ione van enkele valensie die Y-draende sperm aantrek wat lei tot 'n manlike embrio en hoe katione van dubbele valensie die X-draende sperm aantrek wat lei tot 'n vroulike embrio ten spyte daarvan dat dit positief gelaai is. Die studie bevestig verder die natuurlike vroulike oorheersing deur aan te toon dat dit die eierselle en dus by implikasie die vrou is wat die geslag van die embrio bepaal. Die studie bevestig dus uiteindelik dat die dieet ioniese uitwerking op geslagsverhoudings gebruik kan word vir die voorkoming van X-gekoppelde afwykings.

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