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Very Interesting Reading this is the introduction only - see hyperlink below for more: 

UNIVERSITA’ DEGLI STUDI DI NAPOLI FEDERICO II 

DOTTORATO DI RICERCA IN PRODUZIONE E SANITÀ DEGLI ALIMENTI DI ORIGINE ANIMALE INDIRIZZO: SCIENZE DELL’ALLEVAMENTO ANIMALE 

XXIII CICLO 

EFFECT OF SEASON ON REPRODUCTIVE PERFORMANCES IN BUFFALO SPECIES (BUBALUS BUBALIS)

Co-tutor Tutor Chiar.ma Prof.ssa Bianca Gasparrini Chiar.mo Prof. Luigi Zicarelli Coordinatore Chiar.ma Prof.ssa Maria Luisa Cortesi Candidata Dott.ssa Serena Di Francesco

TRIENNIO 2007 - 2010 2

I am strongly inclined to suspect that the most frequent cause of variability may be attributed to the male and female reproductive elements having been affected prior to the act of conception. Charles Darwin, The Origin of Species. 

To My Parents, My Sister, My Friends, To Bianca, Prof Zicarelli, My Colleagues. To Everyone who believed in me and still do. Serena 3

INDEX page INTRODUCTION Mediterranean Italian Buffalo……………………………………… 5 

INTRODUCTION Mediterranean Italian Buffalo

Buffalo is an important economic resource for Italian animal husbandry, especially for the regions traditionally involved in the rearing of this species. The largest concentration of heads can be found in Campania, in the provinces of Caserta and Salerno, where 78% out of 400,000 animals of Italian total amount of buffaloes is bred, but many farms exist also in Latium, in the provinces of Latina and Frosinone, and in Apulia. In these areas, buffalo breeders have constantly grown both in professional and managerial terms, fine-tuning and perfecting breeding techniques and coming to an efficient intensive or semi-intensive approach. In fact, in Italy, the type of farming has changed over time and the scheme with extensive use of meadows and pastures of the past has given way to an intensive farming, with free housing of the animals, modeled on that used for dairy cattle. Dairy buffaloes are kept loose in paddocks close to the milking room, where the cows are submitted to a rigorous control and cleaning of the udder, then mechanically milked twice a day. This advance made the selection of subjects with the purpose of increasing productive and genetic value possible, thus converting a marginal sector into an area with great economic potential. In fact, this breeding has still a positive 6

trend as it is demonstrated by several investments undertaken by many entrepreneurs. Currently, there is a buffalo population growth of about 70,000 animals per year, so the replacement of other species with buffalo is actually performed in many regions. This process is mainly due to the marked productive qualities, adaptability and rustic nature of the buffalo, on one side, but is also promoted –on the other hand- by a lower environmental impact of this species: in fact, it was shown that buffalo eliminates a smaller amount of nitrogen with manure (54 kg vs. 81 kg/year) compared to dairy cow. This allows to increase the load of animals per hectare. Furthermore, in the total budget of a dairy farm, now more than ever, a careful balance between production costs and management costs is necessarily required in order to keep market competitiveness. This, however, must not definitely affect the parameters that allow to obtain a high-quality raw material (milk), because it represents the starting point of a peculiar and strictly linked product, such as mozzarella cheese, which has not only a traditional and cultural value in Italian gastronomy, but above all has been providing employment for many years. In fact, it is worth to highlight that nowadays, all around the world, consumers play a significant role in demanding animal products that are produced to agreed standards for human health, environmental management and animal welfare. Moreover, in the areas mentioned above, the employment impact of this 7

sector is estimated slightly higher than 5%: this can be considered a respectable value, considering that, in Campania, the employment impact in the sectors of agriculture and animal husbandry represents 3.8%, compared to a National value of 2.8% and 1% in Lombardia region.

The importance and competitiveness of buffalo breeding in Italy, compared with the other more established forms of livestock rearing, is also confirmed by the increase in the national buffalo population in the last ten years (+68.64%; source: ANASB1 data, 2008). The buffalo farming in Italy has been reaching remarkable productive standards thanks to an intense work of selection and research carried out during the past years. The most important business related to buffalo breeding is milk production, all of which is processed into mozzarella cheese. The milk average production recorded in 2009 was over 2150 kg with peaks higher than 5000 kg per standard lactation phase (270 days), with 8.39% fat and 4.61% protein content (source: ANASB statistical data, 2009; see Table.1) and with excellent cheese making yields.

1 Associazione Nazionale Allevatori Specie Bufalina, i.e. National Buffalo Breeders' Association. 8

See Full Text Here: http://www.fedoa.unina.it/8229/1/Di_Francesco_Serena_23.pdf

Reproductive biotechnologies………………….............................. 16 

Seasonality of reproduction…………………………....................56 

GAMETES Experiment 1. Assessment of morphological quality and developmental competence to in vitro embryo production (IVEP) of oocytes throughout the seasons……………………………………66 

Materials and methods………………………………………………...67 

Statistical analysis………………………………………………………72 

Results………………………………………………………………......73 Discussion………………………………………………………………77 Experiment 2. Evaluation of some parameters of sperm fertility throughout the seasons……………………………………………84 

Materials and methods…………………………………………………88 

Statistical analysis………………………………………………………90 

Results…………………………………………………………………..90 

Discussion………………………………………………………………93 4 

APPLICATIONS OF INNOVATIVE REPRODUCTIVE BIOTECHNOLOGIES TO BUFFALO BREEDING Experiment 3. 

Artificial insemination (AI) performed in different periods of the year……………………………………………….100 

Materials and methods………………………………………………102 

Statistical analysis……………………………………………………104 

Results…………………………………………………………….......104 

Discussion…………………………………………………………….105 

Experiment 4. Ovum pick-up and in vitro embryo production (OPU - IVEP) trials carried out during transitional period, spring-summer and autumn.…………………………………………………......115

 Materials and methods………………………………………………117 

Statistical analysis……………………………………………………..121 

Results……………………………………………................................121

 Discussion……………………………………………………………..126 CONCLUSIONS……………………………………………………135 REFERENCES………………………………………………140 ACKNOWLEDGMENTS…………………………………..165 5

 

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