Fertility Cryopreservation

The cryobiology is the science of low temperature biology. Fertility cryopreservation is a vital branch of reproductive science and involves the preservation of gametes (sperm and oocytes), embryos, and reproductive tissues (ovarian and testicular tissues) for use in assisted reproduction techniques (ART). The cryopreservation of reproductive cells is the process of freezing, storage, and thawing of spermatozoa or oocytes. It involves an initial exposure to cryoprotectants, cooling to subzero temperature, storage, thawing, and finally, dilution and removal of the cryoprotectants, when used, with a return to a physiological environment that will allow subsequent development. Proper management of the osmotic pressure to avoid damage due to intracellular ice formation is crucial for successful freezing and thawing procedure. Management of non-cryopreserved reproductive cells (i.e., spermatozoa or oocytes) and tissues (i.e., testicular tissue or ovarian tissue) is problematic due to difficulties in donor-recipient synchronization and the potential for transmission of infectious pathogens, which cumulatively limits widespread application of these techniques. Cryopreserved cells and tissues can endure storage for centuries with almost no change in functionality or genetic information, making this storage a method highly attractive. There is a pressing need for the development of optimum cryopreservation methods for reproductive cells and tissues from many species. There are two major techniques for cryopreservation: freeze-thaw processes and vitrification. The major difference between them is the total avoidance of ice formation in vitrification. However, the biotechnology of the reproduction, although widely implemented, has generated protocols currently used to cryopreserve bovine sperm or oocytes, for example, that are still suboptimal, and cannot readily be extrapolated to other species' gametes. ART provide an ensemble of strategies for preserving fertility in patients and commercially valuable or endangered species. Nevertheless, it is very difficult to successfully cryopreserve. Currently, there is a growing interest to understand the underlying cryobiological fundamentals responsible for low survival rates in an effort to develop better cryopreservation. The key factors that affect the life-span of spermatozoa are the combinations of storage temperature, cooling rate, chemical composition of the extender, cryoprotectant concentration, reactive oxygen species (ROS), seminal plasma composition and hygienic control. Sperm preservation protocols vary among animal species owing to their inherent particularities that change extenders used for refrigeration and freezing. On the other hand, oocytes are available only in limited number as compared to spermatozoa, therefore, a cryopreservation protocol must allow a high rate of viability maintenance when they are employed in practical application in ART programs. One of the key factors that influence the freezing process is the ratio of surface area to volume. The oocytes require a longer time to reach osmotic balance with the cryoprotectant solution than the spermatozoa, due to their bigger volume. Then, during cooling of oocytes, various forms of cellular damage may occur, including cytoskeleton disorganization, chromosome and DNA abnormalities, spindle disintegration, plasma membrane disruption and premature cortical granule exocytosis with its related hardening of the zona pellucida. Therefore, animal gametes have been shown to survive storage at low temperatures, and recent results are very encouraging, although reproducible methods have yet to be obtained in many species

Serotonin transporter transgenic (SERTcre) mouse line reveals developmental targets of serotonin specific reuptake inhibitors (SSRIs)

n/

Glutathione concentration and gamma-glutamyltransferase activity in water buffalo colostrum

Evidence is presented that the buffalo mammary gland contains enzymes that catalyse the synthesis and utilization of glutathione. A significant, inverse correlation (r = 0.79) was detected between colostrum gamma-glutamyltransferase (GGT) and glutathione (GSH), suggesting that the enzyme uses GSH as a substrate for its activity. A similar trend was shown in mammary gland homogenates (r = 0.75). Our results show that GSH is secreted into buffalo colostrum and suggest that the enzyme GGT degrades it. To the authors' knowledge, this is the first demonstration of the involvement of GGT-mediated GSH metabolism in the synthesis of colostrums, which elucidates the role of the enzyme that has always been reported very high in colostrum. © 2009 Blackwell Verlag GmbH

The Potential Impact of a Dog Training Program on the Animal Adoptions in an Italian Shelter

SIMPLE SUMMARY: Human–dog relationships are mainly focused on the physical and emotional wellbeing, and has much evolved in the last decades, becoming even more intense over time. Such a novel conception should also be applied to the life that takes place in kennels, which have been mistakenly regarded as a landfill or a burden on society for too many years. Here, we took into consideration an Italian shelter, and analyzed 555 adopted dogs who underwent a well-detailed behavioral training program, to assess the potential impact of the education upon the adoption of attitude. We documented a higher increase in the number of adoptions both for adult and senior animals when compared to the age-matched untrained dogs. Collectively, our data highlight the importance of a proper training, mainly accomplished with reward methods, to get a more suitable and balanced owner-dog attachment. ABSTRACT: One of the main concerns of the human–dog relationship is today associated with the quality life inside the kennels, which are very often regarded as animal dump where dogs are exiled, representing a burden on society. In the present study we sought to investigate the importance of performing an appropriate behavioral program on the adoption chances within an Italian shelter, near Naples (Ottaviano). In this respect, we enrolled 555 adopted dogs of different ages, who followed a tailored-4-month lasting training program between 2018 and 2020. Once entered there, they were carefully examined by the veterinary behaviorist, and directed towards a suited training program, to improve living conditions. We documented a higher number of both adult and senior dogs who left the kennel and were adopted, compared to the age-matched untrained animals (n = 479), housed in the same kennel from 2015 to 2017. Taken together, the present data highlight an important role for training in improving the natural attitudes of the companion dogs, thus pointing towards a better human–animal bond

Combining cannabis and melatonin treatment with a rehabilitation program improved symptoms in a dog with compulsive disorder: A case report

: Compulsive disorder in dogs (CD) is characterized by constant and time-consuming repetition of behaviors, emancipated from the environment, that definitely compromise their everyday life activities. Here, we documented the efficacy of a novel approach to counteract the negative symptoms of CD in a 5-year-old mongrel dog, previously found to be resistant to the conventional antidepressant. The patient underwent an integrated and interdisciplinary approach, based on the cannabis and melatonin co-administration, together with a tailored 5-month-lasting behavioral program. Observational findings showed a lower rate of compulsive episodes and better management of the dog as well, when compared to the previous paroxetine treatment. We followed him for an additional four months of therapy, and the owners reported easier management of the dog, as reduction of abnormal behaviors to a level acceptable to the owners. Overall, our data so far collected in the CD dog may allow us to test more deeply the feasibility and safety of such an off-label approach, at both preclinical and clinical levels

VALUTAZIONE DEL CALCIO INTRACELLULARE IN PIASTINE DI CANI SANI ED AFFETTI DA LEISHMANIOSI

Le piastrine rappresentano gli elementi cellulari fondamentali per il processo di regolazione dell’emostasi. La loro interazione con l’endotelio vasale riveste un ruolo fondamentale sia nella fisiologia che nella fisiopatologia della coagulazione. fisiologicamente, infatti, le piastrine hanno la capacità di aderire alla parete del vaso solo dopo attivazione da parte dell’endotelio vasale. In condizioni patologiche, come quelle causate dalla rottura di una placca aterosclerotica, le piastrine aderiscono all’endotelio, si attivano e reclutano numerose altre piastrine per la formazione del tappo piastrinico. In medicina veterinaria, diversamente dalla medicina umana, la conoscenza dei meccanismi fisiologici specifici alla base di tali processi è piuttosto scarsa. Le piastrine subiscono fenomeni di attivazione, fra i quali è possibile menzionare, lo shape change, l’attivazione del recettore per il fibrinogeno che causa l’aggregazione delle piastrine, il rilascio del contenuto dei granuli e la produzione di trombossano A2 (Gachet C. et al., 1996, Mills D.C.B., 1996). Tali fenomeni sono mediati da modificazioni del calcio intracellulare che origina dal rilascio di calcio dagli store intracellulari e dall’entrata di calcio dal mezzo extracellulare (Rink T.J. et al., 1985, Rink T.J. et al., 1990). D’altro canto però, sono note numerose patologie nel cane, tra cui forme di Leishmaniosi eo Ehrlichiosi, che comportano seria compromissione della funzione coagulativa (Harrus S. et al., 1996; Weiss D.J. et al., 1995). La leishmaniosi, in particolare, è una zoonosi largamente diffusa nelle zone costiere del Mediterraneo caratterizzata da una alterazione della coagulazione spesso asintomatica che si inserisce in un quadro sintomatologico estremamente vario. In precedenti ricerche abbiamo mostrato che, in corso di leishmaniosi, si verifica un danno piastrinico di entità variabile che, peraltro, non trova completa risoluzione con le terapie normalmente utilizzate in corso di tale patologia. I meccanismi attraverso i quali l’aggregazione piastrinica risulta alterata in corso di leishmaniosi non sono ancora chiari. Per tale motivo, l’obiettivo di questo studio è stato quello di valutare le variazioni di calcio nelle piastrine di cani sani ed affetti da leishmaniosi allo scopo di raggiungere una migliore comprensione dei meccanismi responsabili di alterazioni dell’emostasi in corso di tale patologia

Passive transfer status and growth performance in newborn buffalo calves allowed to nurse the dam

The objective of this study was to evaluate the effect of passive transfer status, determined by measuring serum IgG concentration 24 hours after parturition, on growth performance in buffalo calves fed milk replacer or allowed to nurse the dam during the first month of life. Experiment consisted of 24 healthy buffalo calves from birth to 30 days old. Significant quadratic associations were detected between serum IgG concentration 24 hours after birth and day-30 weight (P < 0.05; R2 = 0.62) and between serum IgG concentration 24 hours after birth and the mean daily gain from birth to day 30 (P < 0.01; R2 = 0.74) in buffalo calves allowed to nurse the dam. No significant association was detected between serum IgG concentration 24 hours after birth and measures of growth performance in buffalo calves fed milk replacer. Results indicated that passive transfer status, determined as serum IgG concentration 24 hours after birth, was a significant source of variation in growth performance when buffalo calves nursed the dam. Maximizing passive transfer of immunity by allowing buffalo calves to nurse the dam increased the growth performance during the first month of life

Human-Animal Relationship Dysfunction: A Case Study of Animal Hoarding in Italy

"Animal hoarding" or "compulsive hoarding of animals" is a psychiatric disease, which has important social implications and a profound influence on animal welfare. To date, this phenomenon has been little investigated and largely unexplored. The present study aims to systematically describe a case of animal hoarding, which remains unresolved. The report refers to a case of a woman suffering from animal hoarding that emerged in 2005. From March 2014 to December 2019, 450 animals were seized over nine different occasions. This disease had significant implications on the welfare of the animals collected, which lived in poor housing and hygiene conditions that frequently led to their death. Since animal hoarding cases involve sanitary, legal, and veterinary aspects, we believe that a multidisciplinary approach is necessary in order to prevent a recurrence and a new accumulation of animals. A holistic approach should be taken according to the One Health principle that involves different stakeholders at every level in order to adopt an efficient solution

Expression levels of the focal adhesion-associated proteins paxillin and p130CAS in canine and feline mammary tumors

International audiencePaxillin and p130$^{{\rm CAS}}$ are two adaptor proteins localized at the focal adhesions which play an important role in cell signaling, cell motility and oncogenic transformation. In this study we evaluated the levels of paxillin and p130$^{{\rm CAS}}$ in feline and canine mammary tumor tissues at different stages of malignancy. The results obtained by Western blotting analysis showed no significant differences in the amounts of paxillin and p130$^{{\rm CAS}}$ between normal and non-invasive tumor tissues. By contrast, mammary tumor tissues with the invasive phenotype showed lower levels of paxillin $P < 0.01$ and higher levels of p130$^{{\rm CAS}}$ $P < 0.001$ than normal tissues. The decrease $P < 0.001$ of the amount of paxillin and the increase $P < 0.001$ of p130$^{{\rm CAS}}$ levels were correlated with the progression stage of malignancy. Since paxillin and p130$^{{\rm CAS}}$ are involved in regulating cell migration, our results suggest that low levels of paxillin together with high levels of p130$^{{\rm CAS}}$ expression may cause certain breast cancers to be more motile and possibly more aggressive. Thus, both paxillin and p130$^{{\rm CAS}}$ may represent useful prognosticators of feline and canine breast cancer malignancy

Expression and localization of aquaporin 1 along the intestine of colostrum suckling buffalo calves

Aquaporin 1 (AQP1), a six-transmembrane domain protein, belongs to a highly conserved group of proteins called aquaporins known to regulate permeability across cell membranes. Although the role of AQP1 has been extensively studied, its specific activity along the gastrointestinal tract in animals during early postnatal development is poorly known. This study investigates the expression of AQP1 mRNA and protein in the small and large intestine of water buffalo calves after colostrum ingestion by using reverse transcription polymerase chain reaction (RT-PCR), western blotting, and cellular localization of AQP1 by immunohistochemistry. Our results revealed AQP1 immunoreactivity and the presence of the corresponding mRNA in all the examined tracts of the intestine but with a different cellular localization. Western blotting confirmed the presence of AQP1, with a more intense band in colostrum-suckling animals. These findings offer insights into AQP1 expression in the small and large intestine, suggesting its involvement in osmoregulation in gastrointestinal physiology particularly during the first week after birth in relation to specific maturation of intestinal structures