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in the present study). Only when they reach reproductive age
and veterinary practitioners establish the lack of pregnancy,
this abnormality can be revealed by cytogenetic analysis, PCR
analysis with Y-specific PCR primers, or by FISH mapping
using X/Y chromosome painting probes or bovine/caprine
BAC-clones (Hassaname et al., 1999; Piumi et al., 2001; Di
Berardino et al., 2004). Freemartin condition in both cattle and
river buffalo could be also a very interesting model to study the
development of the immune system and immuno-tolerance, as
well as the fate and potential differentiation of intrauterine cir-
culating stem cells.
In conclusion, all freemartin female river buffaloes were
sterile. This suggests the advisability of their prompt elimina-
lems in male river buffaloes as found in some male cattle co- tion at birth or at early age, especially when karyotype (or
twins of females (Dunn et al., 1979). FISH mapping with appropriate probes, or PCR analysis) con-
Cytogenetic investigations performed in some animals firms this pathology. Eliminating female animals, which will
(cases 1, 2 and 8) (Table 1) in two experiments at different dates never produce calves (and milk), from the farm would lead to
confirmed the same percentage of male and female blood cells. savings in time and money. Sound cooperation among breed-
These results partially agree with those found previously in 22 ers, veterinary practitioners and cytogeneticists is essential if
bulls co-twins (Dunn et al., 1979). Indeed, the latter found we really intend to enhance the genetic improvement of this
some small variations in the frequency of male and female cells species.
of these animals when analyzing the frequency of male and
female cells at different ages.
In river buffalo, since males are normally culled the day
after birth because there is no meat market as yet, at least in
Italy, the breeders found it difficult to cull the females as well,
although their freemartin condition would suggest prompt
elimination from the farm. Male co-twins often die during early
embryonic life and there is no way for breeders to know wheth-
er the female calf born is a freemartin (most cases investigated

Cytogenet Genome Res 108:355-358 (2005) 357