The companys own tests revealed that its wastewater contained many heavy metals in concentrations sufficiently high to bring about serious environmental degradation, including lead, mercury, manganese, arsenic, thallium, and copper, plus the chalcogen selenium. Identifying which particular poison was responsible for the disease proved to be extremely difficult and time-consuming. During 1957 and 1958, many different theories were proposed by different researchers. At first, manganese was thought to be the causal substance due to the high concentrations found in fish and the organs of the deceased. Thallium, selenium, and a multiple contaminant theory were also proposed, but in March 1958, visiting British neurologist Douglas McAlpine suggested that Minamata symptoms resembled those of organic mercury poisoning, so the focus of the investigation centered on mercury. In February 1959, the mercury distribution in Minamata Bay was investigated. The results shocked the researchers involved. Large quantities of mercury were detected in fish, shellfish, and sludge from the bay. The highest concentrations centred around the Chisso factory wastewater canal in Hyakken Harbour and decreased going out to sea, clearly identifying the plant as the source of contamination. Pollution was so heavy at the mouth of the wastewater canal, a figure of 2 kg of mercury per ton of sediment was measured: a level that would be economically viable to mine. Indeed, Chisso did later set up a subsidiary to reclaim and sell the mercury recovered from the sludge.Hair samples were taken from the individuals with the disease and also from the Minamata population in general.
Outbreak of Niigata Minamata disease
Minamata disease broke out again in 1965, this time along the banks of the Agano River in Niigata Prefecture. The polluting factory (owned by Showa Denko) employed a chemical process using a mercury catalyst very similar to that used by Chisso in Minamata. As in Minamata, from the autumn of 1964 to the spring of 1965, cats living along the banks of the Agano River had been seen to go mad and die. Before long, patients appeared with identical symptoms to patients living on the Shiranui Sea, and the outbreak was made public on 12 June 1965. Researchers from the Kumamoto University Research Group and Hajime Hosokawa (who had retired from Chisso in 1962) used their experience from Minamata and applied it to the Niigata outbreak. In September 1966, a report was issued proving Showa Denkos pollution to be the cause of this second Minamata disease outbreak. Unlike the patients in Minamata, the victims of Showa Denkos pollution lived a considerable distance from the factory and had no particular link to the company. As a result, the local community was much more supportive of patients groups and a lawsuit was filed against Showa Denko in March 1968, only three years after discovery. The events in Niigata catalysed a change in response to the original Minamata incident. The scientific research carried out in Niigata forced a re-examination of that done in Minamata and the decision of Niigata patients to sue the polluting company allowed the same response to be considered in Minamata.