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.