**9. Transmission of reproductive diseases**

The methods used for assisted reproduction in wild animals should be free of contamination or diseases, therefore, some measures must be taken to avoid compromising the procedures. The semen centrifugation and seminal plasma removal are essential for artificial insemination with fresh or frozen semen and *in vitro* fertilization procedures.

treatment with exogenous gonadotropins and laparoscopy are the basic requirements for

The immature oocytes collected ( 60%) in cats become mature in 24-32 hours in culture media. These about 70% are fertilized, however, only a small percentage, from 20 to 30%

Donoghue et al. (1990) reported the birth of the first wild cat from IVF after an embryo transfer. Tiger cubs (*Panthera tigris*) were produced *in vitro* using excellent quality embryos, containing two to four cells, and surgically transferred to the oviducts of two females. Pregancy was successful in one of these females and three kittens were born after 107 days. The first embryos produced by IVF in Brazil were jaguar (*Panthera onca*) embryos. The ovarian stimulation with pFSH/LH produced 25 follicles/female ( 80%), however, despite the recovery of high quality oocytes, the fertilization percentages were low ( 25%)

In a Project involving the São Paulo University/Brazil (USP), the Mata Ciliar Association/Brazil (AMC) and the Cincinnati Zoo/USA (CREW), 128 ocelot (*Leopardus pardalis*) and tigrina (*Leopardus tigrinus*) embryos were produced. The animals were treated with eCG/hCG and produced 10 follicles/female. Follicles were aspirated by laparoscopy and 7-9 of excellent quality oocytes/female were recovered. Of these, 60% were fertilized *in vitro*, resulting in 76 ocelot and 52 tigrina embryos (Swanson & Brown, 2004). Two ocelots became pregnant with normal pregnancy development and birth, one from the Cincinnati Zoo/USA and another from the Sao Bernardo do Campo Zoo/Brazil. However, only the

Among all the Neotropical cats, the ocelot is the only species that produced offspring after the transfer of frozen embryos (Swanson, 2001, 2002b). These births were the result of a cooperative effort between Brazil and the USA for the development of adequate management programs for this species in captivity, and for the establishment of genetically

Based on the percentage of successful embryonic cleavage after thawing domestic cat embryos, which is less than 70% (Pope, 2000), the difficulties to develop feline embryos used post-freezing is recognized. The difficulties may be related to inappropriate timing for the embryo implantation or fetal survivel (Swanson & Brown, 2004) and not only related to the quality of the embryos. Thus, it is necessary to detect the female natural receptor estrus in order to perform the transfer of thawed embryos. This determination is achieved through measurement of fecal steroid levels according to the enzyme-immunoassay (EIA) technique

After the determination of the natural estrus, the ovulation should be induced with GnRH (Gonadotropin Releasing Hormone) and the embryos should be transferred by video-

The methods used for assisted reproduction in wild animals should be free of contamination or diseases, therefore, some measures must be taken to avoid compromising the procedures. The semen centrifugation and seminal plasma removal are essential for artificial

insemination with fresh or frozen semen and *in vitro* fertilization procedures.

offspring born to the American female survived after birth (Swanson, 2002b).

viable populations between the Brazilian and American populations.

ovarian stimulation and oocyte retrieval for IVF procedures.

develops in blastocysts (Johnston et al., 1989).

(Morato et al., 2000).

for the fecal estrogens metabolites.

laparoscopy directly in the ostium of the oviduct.

**9. Transmission of reproductive diseases** 

The methods for processing semen without centrifugation and removal of the seminal plasma were responsible for the development of pyometra in 40% of inseminated domestic cats, regardless of the semen being diluted in culture medium containing antibiotics penicillin and streptomycin (Howard, 1993). It is believed that the donors might carry bacteria in their normal flora, eg *E. coli*, which would cause infection in the females.

In a study conducted by Paz et al. (1999), aiming at determining preputial microbiota in nine adult male jaguars (*Panthera onca*), the most frequently observed microorganism were *Staphylococcus sp* (40%), followed by *Streptococcus sp* (30%), *Escherichia coli* (20%) and *Corynebacterium sp* (10%).

The preputial microbiota in the genus Leopardus was assessed by Guido et al. (2000), and the results were *Escherichia coli* (40%), *Proteus rettgeri* (40%) and *Yersinia psedotuberculosis* (20%) in tigrina (*Leopardus tigrinus*) (n = 5), *Staphylococcus sp* (42.9%), *Escherichia coli* (28.5%), *Streptococcus sp* (14.3%), *Staphylococcus sp* + *Streptococcus sp* (14.3%) in ocelots (*Leopardus pardalis*) (n = 6) and only *Staphylococcus sp* in margay (*Leopardus wieddi*) (n = 1).

The feline immunodeficiency virus is present in the semen of domestic cats and can be transmitted to females by AI (Jordan et al., 1995, 1996). This aspect should be taken into consideration during assisted reproduction procedures performed in wild cats.

In addition, a reproductive evaluation and clinical examination should be performed in wild cats before the animal inclusion in the management programs using assisted reproduction.
