Artificial insemination (AI) can undoubtedly be regarded as the oldest and most widely used assisted reproductive technique/technology (ART) applied in livestock production and it is one of the most important ARTs. The three cornerstones of its application are that it is simple, economical and successful. Artificial insemination offers many well-known benefits for producers. Fresh, fresh + diluted + chilled and frozen semen can be used for AI in small ruminants. To ensure its successful use, the AI technique must be selected on the basis of the type of semen planned to be used. This review paper gives a detailed overview of semen processing and its effects on semen quality, as well as of the AI techniques applied in small ruminants and their success rates.

Q fever is an important zoonotic disease caused by Coxiella burnetii. There are few reliable data about C. burnetii infection available. The aim of this study was to assess the importance and potential infectious sources of Q fever in Hungary. A total of 215 milk samples (10 individual samples from each herd and 1 bulk tank milk sample from each cattle herd), and 400 serum samples (20 from each herd) were tested from 15 dairy cattle herds and 5 sheep flocks located in different parts of Hungary. The study found 19.3% (58/300) and 38.0% (57/150) seropositivity in cattle, and 0% (0/100) and 6.0% (3/50) seropositivity in sheep, by complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA), respectively. C. burnetii DNA was detected by IS1111 element-based TaqMan real-time polymerase chain reaction (PCR) in 8.7% (13/150) of individual dairy cow milk samples, 4.0% (2/50) of individual sheep milk samples, and 66.7% (10/15) of dairy bulk tank milk samples. Samples taken from nine different commercially-available pasteurized cow milk products from different Hungarian producers were also tested for the presence of C. burnetii DNA, and eight of these samples were found to be positive (88.9%). The real-time PCR examination of 5402 ixodid ticks collected from different parts of the country yielded negative results. Knowledge of the true prevalence of Q fever is crucial for policymakers involved in evidence-based decision making.

By decreasing the volume of the cryoprotective solution it is possible to increase dramatically the freezing speed and -at the same time -reduce the toxicity and osmotic side effects of cryoprotectants (CPA). The objective of our study was to vitrify Day-3 cleavage stage mouse embryos (n = 229) with the cryoloop technology using a new composition of vitrification media. Embryos were exposed to a 2-step loading of CPA, ethylene glycol (EG) and propylene glycol (PG), before being placed on the surface of a thin filmy layer formed from the vitrification solution in a small nylon loop, then they were rapidly submerged into liquid nitrogen. After warming, the CPA was diluted out from the embryos by a 3-step procedure. Survival of embryos was based on morphological appearance after thawing and continued development to expanded blastocysts upon subsequent 48-hour culture. Embryos of the two control groups were either treated likewise except that they were not vitrified, or cultured in vitro without any treatment. Our data show that a high percentage of embryos survived (92.7%) vitrification in the mixture of EG and PG combined with cryoloop carrier and developed normally (89.1%) in vitro after thawing. To our knowledge this is the first report of the successful vitrification of cleavage stage mouse embryos using VitroLoop vitrification procedure.

In this study, we present an optimised colourimetric and a lateral flow LAMP assay for the detection of Haemonchus contortus in small ruminant faecal samples. Using a previously published LAMP primer set, we made use of commercially available colourimetric LAMP and lateral flow kits and combined this into an optimised diagnostic assay which was then tested on field faecal samples from Eastern and South-Eastern Hungary as well as a pure H. contortus egg faecal sample from Košice, Slovakia. Both assays showed no conflicts in visual detection of the results. Additionally, we modified and tested several centrifuge-free DNA extraction methods and one bead-beating egg lysis DNA extraction method to develop a true point of care protocol, as the source of the starting DNA is the main rate-limiting step in farm-level molecular diagnosis. Out of the various methods trialed, promising results were obtained with the magnetic bead extraction method. Sample solutions from the Fill-FLOTAC® technique were also utilised, which demonstrated that it could be efficiently adapted for field-level egg concentration to extract DNA. This proof of concept study showed that isothermal amplification technologies with a colourimetric detection or when combined with a lateral flow assay could be an important step for a true point of care molecular diagnostic assay for H. contortus.

The complex anatomical structure of the ewe reproductive tract accompanied with low quality of frozen ram semen for artificial insemination, resulted in a challenge with regard to using superior genotypes for reproductive ovine performance. Hence, improved genetics in ovine management has not been efficiently and widely used especially in undeveloped countries. Therefore, intrauterine semen deposition by laparoscopic insemination should be adopted in the current sheep production systems. Thus, this study aimed to assess the pregnancy rate and lambing rate of ewe inseminated by laparoscopic insemination techniques using frozen-thawed semen. The research used imported frozen semen from two rams of the Lacaune breed. Ewes were grouped according to age in years (1, 2 and 4). Before insemination, the semen was examined microscopically for its motility and viability and thereafter the laparoscopic artificial insemination technique was performed to 19 Lacaune breed ewes using frozen-thawed semen. The overall pregnancy and prolificacy rates were 31.57% and 42.10% respectively. Out of 2 ewes in the 1-year age group that were inseminated, only 1 ewe lambed representing 50%. However, from 16 ewes inseminated of four-year age group, 5 ewes lambed representing 31.25%. Significant difference based on age group was not evaluated due disproportionate of the data, (such that the data included 2 ewes in one-year-old age, 1 ewe in 2-year-old age and 16 ewes in 4-year-old age). Based on the ram semen, 33.33% and 30% of the inseminated ewes were pregnant from ram A and ram B semen respectively. However, in the case of prolificacy rate, 44.44% and 40 % of the ewes lambed from using semen of ram A and B, respectively. There was no significant difference (p>0.05) in pregnancy and prolificacy rates based on semen from the two rams. In conclusion, in this research study, ram semen had no significant effect on pregnancy and prolificacy rates using laparoscopic AI on Lacaune sheep. This could be due to the fact that the rams had very good quality semen. Evaluation of ram semen, accompanied with appropriate ewe selection based on age and rightful deposition of semen could lead to better and more consistent results. Overall this could contribute to the successful application of laparoscopic artificial insemination in Lacaune sheep production systems for enhanced productivity.

The viewpoint of the recent cryopreservation techniques (CT) suggests the use of a reduced volume of cryopreservation solution, high concentration of cryoprotectants and ultra-rapid cooling and warming rates help to reduce cryo-injury and maximize the viability of the preserved animal genetic resources (AnGR). The CT had now become widely accepted as one of the best methods of choice for the ex-situ conservation of AnGR due to its high success rate recorded and no-invasive nature as compared to the conventional slow rate freezing (CSRF). Rapid advances and wide acceptability of the use of assisted reproductive technologies (ART’s) particularly artificial insemination (AI) in animal breeding had resulted in a greater loss of a large number of good quality genes in virtually almost all the native breeds of animals across the globe. Small ruminant (SR) animals are not an exception in such present predicaments situation of erosion and dilution of the valuable AnGR among the native breeds. As a result of this, 148 and 16 breeds of sheep and goats respectively have already become extinct in Europe and the Caucasus. In view of the aforementioned situation, the present review aimed at exploring some of the current states of development, roles played and potentials of CT in the conservation of SR genes and genome for the immediate and future breeding purposes for sustainable development. It basically covers; animal genetic resource, the need to conserve AnGR, tools for ex situ in vitro conservation of AnGR and recent developments in breeding and cryopreservation of SR AnGR. Cryopreservation is playing a pivotal role in ex-situ gene conservation of AnGR. Decline in genetic diversity among SR breed population was high in Europe and the Caucasus. There is therefore, need for improvent on current stringent measures on conservation of AnGR in this region of the world.

The semen of domestic mammals is conventionally collected with an artificial vagina (AV) for artificial insemination (AI) or for short- or long-term storage. However, the procedure has certain drawbacks: animal training is not feasible in extensive animal care systems nor among wild species, as the trained animals sometimes fail to mount. Hence, there is a need for alternative semen collection methods. Electroejaculation (EEJ) and epididymal sperm recovery (ESR) are the two effective alternatives to AV. However, in recent years, animal welfare campaigners have called for the ban, in certain EU countries, of EEJ due to its inhumane nature. In this review, alternative methods of sperm collection (by EEJ and ESR, their qualities, and their freezing techniques) are highlighted, as well as the effects of EEJ on pre-freeze and post-thaw ram sperm quality parameters and the animal welfare progress made in EEJ between the 20th and 21st centuries. Additionally, the techniques for enhancing post-thaw sperm quality prior to freezing and for the freezing of EEJ and ESR spermatozoa are explored. ESR and EEJ are reliable alternatives to AV on certain occasions. EEJ is ideal for semen collection in wild or untrained animals, breeding soundness examinations, collection outside of the breeding season, and culling. At the same time, ESR is ideal in cases of castration, accidental death of elite sire, or postmortem for gene conservation purposes or assisted reproductive technologies (ARTs) studies.

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